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Hanamura T, Christenson JL, O'Neill KI, Rosas E, Spoelstra NS, Williams MM, Richer JK. Secreted indicators of androgen receptor activity in breast cancer pre-clinical models. Breast Cancer Res 2021; 23:102. [PMID: 34736512 PMCID: PMC8567567 DOI: 10.1186/s13058-021-01478-9] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/06/2021] [Accepted: 10/19/2021] [Indexed: 01/01/2023] Open
Abstract
PURPOSE Accumulating evidence has attracted attention to the androgen receptor (AR) as a biomarker and therapeutic target in breast cancer. We hypothesized that AR activity within the tumor has clinical implications and investigated whether androgen responsive serum factors might serve as a minimally invasive indicator of tumor AR activity. METHODS Based on a comprehensive gene expression analysis of an AR-positive, triple negative breast cancer patient-derived xenograft (PDX) model, 163 dihydrotestosterone (DHT)-responsive genes were defined as an androgen responsive gene set. Among them, we focused on genes that were DHT-responsive that encode secreted proteins, namely KLK3, AZGP1 and PIP, that encode the secreted factors prostate specific antigen (PSA), zinc-alpha-2-glycoprotein (ZAG) and prolactin induced protein (PIP), respectively. Using AR-positive breast cancer cell lines representing all breast cancer subtypes, expression of candidate factors was assessed in response to agonist DHT and antagonist enzalutamide. Gene set enrichment analysis (GSEA) was performed on publically available gene expression datasets from breast cancer patients to analyze the relationship between genes encoding the secreted factors and other androgen responsive gene sets in each breast cancer subtype. RESULTS Anti-androgen treatment decreased proliferation in all cell lines tested representing various tumor subtypes. Expression of the secreted factors was regulated by AR activation in the majority of breast cancer cell lines. In GSEA, the candidate genes were positively correlated with an androgen responsive gene set across breast cancer subtypes. CONCLUSION KLK3, AZGP1 and PIP are AR regulated and reflect tumor AR activity. Further investigations are needed to examine the potential efficacy of these factors as serum biomarkers.
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Affiliation(s)
- Toru Hanamura
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA
| | - Jessica L Christenson
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA
| | - Kathleen I O'Neill
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA
| | - Emmanuel Rosas
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA
| | - Nicole S Spoelstra
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA
| | - Michelle M Williams
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA
| | - Jennifer K Richer
- Department of Pathology, University of Colorado, Anschutz Medical Campus, 12800 E. 19th Ave., Aurora, CO, 80045, USA.
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Xu L, Yu W, Niu M, Zheng C, Qu B, Li Y, Wang J, Huang P, Wang O, Gong F. Serum ZAG Levels Were Associated with eGFR Mild Decrease in T2DM Patients with Diabetic Nephropathy. Int J Endocrinol 2017; 2017:5372625. [PMID: 28352283 PMCID: PMC5352973 DOI: 10.1155/2017/5372625] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/24/2016] [Revised: 12/07/2016] [Accepted: 12/26/2016] [Indexed: 12/26/2022] Open
Abstract
Objective. To investigate the changes of serum zinc-α2-glycoprotein (ZAG) in type 2 diabetes mellitus (T2DM) with eGFR mild decrease. Subjects and Methods. A total of 438 T2DM patients (61.3 ± 4.0 y) were recruited and the demographic, anthropometric, and biochemical parameters were all collected. Serum ZAG levels were determined by commercially available ELISA kits. Results. The proportion of T2DM patients with the high tertile ZAG levels was 11.9% higher in patients with mildly decreased estimated glomerular filtration rate (eGFR) (<90 mL/min/1.73 m2) than those with the low tertile ZAG levels (P = 0.038). The probability of the eGFR < 90 mL/min/1.73 m2 in patients with the high ZAG levels was 94% higher than those with the low serum ZAG levels after adjusting for age, gender, and education [OR = 1.94, 95% CI (1.17-3.23), P = 0.0094]. This phenomenon was more likely to be observed in the condition of uACR ≥ 2.7 mg/mmol, WC ≥ 90 cm for men, or WC ≥ 85 cm for women. Conclusion. Serum ZAG levels were firstly found to be related with eGFR in T2DM patients. The patients with the high tertile ZAG levels were more likely to have mildly eGFR decrease, especially for female patients with higher uACR and bigger WC.
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Affiliation(s)
- Lingling Xu
- Key Laboratory of Endocrinology of National Health and Family Planning Commission, Department of Endocrinology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100730, China
| | - Weihong Yu
- Department of Ophthalmology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100730, China
| | - Meng Niu
- Department of Endocrinology, Traditional Chinese Medicine Hospital of Muping District of Yantai City, Yantai, Shandong, China
| | - Caixia Zheng
- Department of Ophthalmology, Traditional Chinese Medicine Hospital of Muping District of Yantai City, Yantai, Shandong, China
| | - Bin Qu
- Department of Ophthalmology, Traditional Chinese Medicine Hospital of Muping District of Yantai City, Yantai, Shandong, China
| | - Yan Li
- Department of Ophthalmology, Traditional Chinese Medicine Hospital of Muping District of Yantai City, Yantai, Shandong, China
| | - Jing Wang
- Department of Ophthalmology, Traditional Chinese Medicine Hospital of Muping District of Yantai City, Yantai, Shandong, China
| | - Ping Huang
- Department of Ophthalmology, Traditional Chinese Medicine Hospital of Muping District of Yantai City, Yantai, Shandong, China
| | - O. Wang
- Key Laboratory of Endocrinology of National Health and Family Planning Commission, Department of Endocrinology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100730, China
| | - Fengying Gong
- Key Laboratory of Endocrinology of National Health and Family Planning Commission, Department of Endocrinology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100730, China
- *Fengying Gong:
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Yang M, Liu R, Li S, Luo Y, Zhang Y, Zhang L, Liu D, Wang Y, Xiong Z, Boden G, Chen S, Li L, Yang G. Zinc-α2-glycoprotein is associated with insulin resistance in humans and is regulated by hyperglycemia, hyperinsulinemia, or liraglutide administration: cross-sectional and interventional studies in normal subjects, insulin-resistant subjects, and subjects with newly diagnosed diabetes. Diabetes Care 2013; 36:1074-1082. [PMID: 23275352 PMCID: PMC3631846 DOI: 10.2337/dc12-0940] [Citation(s) in RCA: 89] [Impact Index Per Article: 7.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/15/2012] [Accepted: 10/10/2012] [Indexed: 02/03/2023]
Abstract
OBJECTIVE Zinc-α2-glycoprotein (ZAG) has been proposed to play a role in the pathogenesis of insulin resistance. Previous studies in humans and in rodents have produced conflicting results regarding the link between ZAG and insulin resistance. The objective of this study was to examine the relationships between ZAG and insulin resistance in cross-sectional and interventional studies. RESEARCH DESIGN AND METHODS Serum ZAG (determined with ELISA) was compared with various parameters related to insulin resistance in subjects with normal glucose tolerance, impaired glucose tolerance (IGT), and newly diagnosed type 2 diabetes mellitus (T2DM), and in women with or without polycystic ovary syndrome (PCOS). Euglycemic-hyperinsulinemic clamps were performed in healthy and PCOS women. Real-time RT-PCR and Western blotting were used to assess mRNA and protein expression of ZAG. The effect of a glucagon-like peptide-1 agonist on ZAG was studied in a 12-week liraglutide treatment trial. RESULTS Circulating ZAG was lower in patients with IGT and newly diagnosed T2DM than in controls. Circulating ZAG correlated positively with HDL cholesterol and adiponectin, and correlated inversely with BMI, waist-to-hip ratio, body fat percentage, triglycerides, fasting blood glucose, fasting insulin, HbA1c, and homeostasis model assessment of insulin resistance (HOMA-IR). On multivariate analysis, ZAG was independently associated with BMI, HOMA-IR, and adiponectin. ZAG mRNA and protein were decreased in adipose tissue of T2DM patients. Moreover, circulating ZAG levels were lower in women with PCOS than in women with high insulin sensitivity. Liraglutide treatment for 12 weeks significantly increased circulating ZAG levels. CONCLUSIONS We conclude that ZAG may be an adipokine associated with insulin resistance.
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Affiliation(s)
- Mengliu Yang
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Rui Liu
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Shu Li
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Yu Luo
- Key Laboratory of Laboratory Medical Diagnostics in the Ministry of Education and Department of Clinical Biochemistry, College of Laboratory Medicine, Chongqing Medical University, Chongqing, China
| | - Yali Zhang
- Key Laboratory of Laboratory Medical Diagnostics in the Ministry of Education and Department of Clinical Biochemistry, College of Laboratory Medicine, Chongqing Medical University, Chongqing, China
| | - Lili Zhang
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Dongfang Liu
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Yaxu Wang
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Zhengai Xiong
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Guenther Boden
- Division of Endocrinology/Diabetes/Metabolism and the Clinical Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania
| | - Shirong Chen
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Ling Li
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
| | - Gangyi Yang
- Department of Endocrinology, Department of Osteology, Department of Obstetrics and Gynecology, Department of Surgery, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
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Kushnir MM, Naessén T, Wanggren K, Rockwood AL, Crockett DK, Bergquist J. Protein and steroid profiles in follicular fluid after ovarian hyperstimulation as potential biomarkers of IVF outcome. J Proteome Res 2012; 11:5090-100. [PMID: 22988950 DOI: 10.1021/pr300535g] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Controlled ovarian hyperstimulation is performed to assist with generation of multiple mature oocytes for use in in vitro fertilization (IVF). The goal of our study was to evaluate differences in protein and steroid profiles in ovarian follicular fluid (hFF) samples obtained during oocyte retrieval from women undergoing IVF treatment and to identify physiological pathways associated with the proteins. The hFF samples were depleted of abundant proteins, fractionated by ultrafiltration, digested, and analyzed by nano-LC-QTOF. Concentrations of 15 endogenous steroids were determined in the samples using LC-MS/MS methods. The total number of proteins identified in the samples was 75, of which 4, 7, and 2 were unique to the samples from women with viable pregnancy, miscarriage, and no pregnancy, respectively. Identified proteins were associated with the acute response signaling, coagulation system, intrinsic and extrinsic prothrombin activation, complement system, neuroprotective role of THOP1, FXR/RXR activation, role of tissue factor, and growth hormone pathways. A greater number of proteins associated with biosynthesis was found in hFF samples corresponding to the oocytes resulting in pregnancy. The abundance of seven proteins was found to be associated with steroidogenesis. The obtained data will contribute to better understanding of the pathogenesis and development of noninvasive markers for assessment of oocytes viability.
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Affiliation(s)
- Mark M Kushnir
- ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah 84108, United States.
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McDermott L, Jadoon A, Cunningham P. ZAG and a potential role in systemic lipid homeostastis: examining the evidence from in vitro human studies and patients with chronic illness. ACTA ACUST UNITED AC 2012. [DOI: 10.2217/clp.12.45] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022]
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Pitteri SJ, Hanash SM, Aragaki A, Amon LM, Chen L, Busald Buson T, Paczesny S, Katayama H, Wang H, Johnson MM, Zhang Q, McIntosh M, Wang P, Kooperberg C, Rossouw JE, Jackson RD, Manson JE, Hsia J, Liu S, Martin L, Prentice RL. Postmenopausal estrogen and progestin effects on the serum proteome. Genome Med 2009; 1:121. [PMID: 20034393 PMCID: PMC2808737 DOI: 10.1186/gm121] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2009] [Revised: 11/07/2009] [Accepted: 12/24/2009] [Indexed: 12/29/2022] Open
Abstract
Background Women's Health Initiative randomized trials of postmenopausal hormone therapy reported intervention effects on several clinical outcomes, with some important differences between estrogen alone and estrogen plus progestin. The biologic mechanisms underlying these effects, and these differences, have yet to be fully elucidated. Methods Baseline serum samples were compared with samples drawn 1 year later for 50 women assigned to active hormone therapy in both the estrogen-plus-progestin and estrogen-alone randomized trials, by applying an in-depth proteomic discovery platform to serum pools from 10 women per pool. Results In total, 378 proteins were quantified in two or more of the 10 pooled serum comparisons, by using strict identification criteria. Of these, 169 (44.7%) showed evidence (nominal P < 0.05) of change in concentration between baseline and 1 year for one or both of estrogen-plus-progestin and estrogen-alone groups. Quantitative changes were highly correlated between the two hormone-therapy preparations. A total of 98 proteins had false discovery rates < 0.05 for change with estrogen plus progestin, compared with 94 for estrogen alone. Of these, 84 had false discovery rates <0.05 for both preparations. The observed changes included multiple proteins relevant to coagulation, inflammation, immune response, metabolism, cell adhesion, growth factors, and osteogenesis. Evidence of differential changes also was noted between the hormone preparations, with the strongest evidence in growth factor and inflammation pathways. Conclusions Serum proteomic analyses yielded a large number of proteins similarly affected by estrogen plus progestin and by estrogen alone and identified some proteins and pathways that appear to be differentially affected between the two hormone preparations; this may explain their distinct clinical effects.
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Affiliation(s)
- Sharon J Pitteri
- Public Health Sciences Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N, Seattle, WA 98109, USA
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Ceperuelo-Mallafré V, Näf S, Escoté X, Caubet E, Gomez JM, Miranda M, Chacon MR, Gonzalez-Clemente JM, Gallart L, Gutierrez C, Vendrell J. Circulating and adipose tissue gene expression of zinc-alpha2-glycoprotein in obesity: its relationship with adipokine and lipolytic gene markers in subcutaneous and visceral fat. J Clin Endocrinol Metab 2009; 94:5062-9. [PMID: 19846741 DOI: 10.1210/jc.2009-0764] [Citation(s) in RCA: 65] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
Abstract
CONTEXT Zinc-alpha2-glycoprotein (ZAG) is a soluble protein similar to the class I major histocompatibility complex heavy chain, which has been implicated in lipid catabolism. We hypothesized that ZAG mRNA expression in adipose tissue may be linked with lipolytic and adipokine gene expression and have a close relationship with clinical phenotype. OBJECTIVES The objective of the study was to analyze ZAG gene expression in human adipose tissue from lean and obese subjects. ZAG circulating plasma levels and its relationship with cardiometabolic risk factors were also studied. DESIGN Seventy-three Caucasian (43 male and 30 female) subjects were included. Plasma and adipose tissue [sc (SAT) and visceral (VAT)] from the same patient were studied. mRNA of PPARgamma, hormone-sensitive lipase (HSL), adipose triglyceride lipase, adiponectin, omentin, visfatin, and ZAG were quantified. Plasma concentrations of ZAG were determined with ELISA. RESULTS ZAG plasma levels showed a negative correlation with insulin (r = -0.39; P = 0.008) and the homeostasis model assessment for insulin resistance index (r = -0.36; P = 0.016). No differences in ZAG circulating levels according to body mass index classification were observed. ZAG expression in SAT was significantly reduced in overweight and obese individuals compared with lean subjects (P < 0.001 and P = 0.007, respectively). ZAG mRNA expression in both SAT and VAT depots were negatively correlated with many clinical and metabolic cardiovascular risk factors. After multiple linear regression analysis, SAT ZAG was mainly predicted by adiponectin mRNA expression (B = 0.993; P < 0.0001) and plasma triglyceride levels (B = -0.565; P = 0.006). VAT ZAG expression was predicted by adiponectin expression (B = 0.449; P < 0.0001), and HSL VAT expression (B = 0.180; P = 0.023). CONCLUSIONS The present study provides evidence of a role of ZAG gene in adipose tissue metabolism, with a close association with adiponectin gene expression in sc and visceral fat.
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Affiliation(s)
- V Ceperuelo-Mallafré
- Endocrinology and Nutrition Unit, Research Department, University Hospital of Tarragona Joan XXIII, Pere Virgili Institute, 43007 Tarragona, Spain
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Selva DM, Lecube A, Hernández C, Baena JA, Fort JM, Simó R. Lower zinc-alpha2-glycoprotein production by adipose tissue and liver in obese patients unrelated to insulin resistance. J Clin Endocrinol Metab 2009; 94:4499-507. [PMID: 19622624 DOI: 10.1210/jc.2009-0758] [Citation(s) in RCA: 79] [Impact Index Per Article: 4.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
Abstract
CONTEXT Zinc-alpha2 glycoprotein (ZAG) has been proposed as a new candidate in the pathogenesis of obesity, but most of the information stems from studies performed in rodents and in vitro assays. OBJECTIVE The main aim of the study was to compare serum levels of ZAG and its expression (mRNA levels and protein) in adipose tissue and the liver between obese and nonobese subjects. The relationship between ZAG and insulin resistance was also explored. DESIGN This was a case-control study. SETTING The study was conducted at a university referral center. PATIENTS AND METHODS Samples of serum, sc adipose tissue (SAT), visceral adipose tissue (VAT), and liver were obtained from 20 obese subjects during bariatric surgery. Samples from 10 nonobese patients matched by age and gender were used as a control group. Serum ZAG levels were determined by ELISA. ZAG mRNA levels were measured by real-time PCR and protein content by Western blot. The effect of insulin on liver production of ZAG was assessed using HepG2 cultures. RESULTS Serum concentration of ZAG (micrograms per milliliter) was significantly lower in obese subjects (40.87 +/- 10.45 vs. 63.26 +/- 16.40; P = 0.002). ZAG expression was significantly lower in the adipose tissue (SAT and VAT) and liver of obese patients than in control subjects. Significant negative correlations between body mass index and circulating ZAG (r = -0.65, P < 0.001) as well as between body mass index and mRNA ZAG levels in SAT (r = -0.68, P < 0.001) and VAT were detected (r = -0.64, P < 0.001). No relationship was found between ZAG and homeostasis model assessment for insulin resistance and insulin had no effect on ZAG production in vitro. CONCLUSION A down-regulation of ZAG in SAT, VAT, and liver exists in obese patients but seems unrelated to insulin resistance.
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Affiliation(s)
- David M Selva
- Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas, Instituto de Salud Carlos III, Universitat Autònoma de Barcelona, Barcelona, Spain
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Hassan MI, Waheed A, Yadav S, Singh TP, Ahmad F. Zinc alpha 2-glycoprotein: a multidisciplinary protein. Mol Cancer Res 2008; 6:892-906. [PMID: 18567794 DOI: 10.1158/1541-7786.mcr-07-2195] [Citation(s) in RCA: 179] [Impact Index Per Article: 10.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Zinc alpha 2-glycoprotein (ZAG) is a protein of interest because of its ability to play many important functions in the human body, including fertilization and lipid mobilization. After the discovery of this molecule, during the last 5 decades, various studies have been documented on its structure and functions, but still, it is considered as a protein with an unknown function. Its expression is regulated by glucocorticoids. Due to its high sequence homology with lipid-mobilizing factor and high expression in cancer cachexia, it is considered as a novel adipokine. On the other hand, structural organization and fold is similar to MHC class I antigen-presenting molecule; hence, ZAG may have a role in the expression of the immune response. The function of ZAG under physiologic and cancerous conditions remains mysterious but is considered as a tumor biomarker for various carcinomas. There are several unrelated functions that are attributed to ZAG, such as RNase activity, regulation of melanin production, hindering tumor proliferation, and transport of nephritic by-products. This article deals with the discussion of the major aspects of ZAG from its gene structure to function and metabolism.
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Affiliation(s)
- Md Imtaiyaz Hassan
- Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi 110025, India
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Hassan MI, Kumar V, Kashav T, Alam N, Singh TP, Yadav S. Proteomic approach for purification of seminal plasma proteins involved in tumor proliferation. J Sep Sci 2007; 30:1979-88. [PMID: 17638362 DOI: 10.1002/jssc.200700028] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
Human seminal plasma contains a large array of proteins required for the normal physiology and metabolism of spermatozoa. These are mainly secreted from prostate epithelium, testes, and seminal vesicles. Fortunately, many of these are found to be present at elevated concentration in seminal plasma and act as a biomarker of different carcinomas as their levels are also enhanced in serum and are found to be involved in tumor progression and metastasis apart from fertility. The proteins which were overexpressed in the seminal plasma of prostate carcinoma patients were identified by 2-DE and MALDI-TOF/MS. We have designed a strategy to purify these four proteins prostate specific antigen (PSA), prostatic acid phosphatase (PAP), Zinc alpha2-glycoprotein (ZAG), and progastricsin (PG), together in homogeneity by using simple chromatographic techniques. Acidic and basic fractions of human seminal plasma were separated by ion exchange chromatography and further purified by gel permeation chromatography. Our results form a new and valuable resource for those attempting structure-based drug designing for prostate and other cancers where the amount of proteins is required in plenty and in native form.
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Affiliation(s)
- Md Imtaiyaz Hassan
- Department of Biophysics, All India Institute of Medical Sciences, New Delhi 110029, India
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11
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Gonzalez LO, Corte MD, Junquera S, Bongera M, Rodriguez JC, Vizoso FJ. Expression of androgen receptor and two androgen-induced proteins (apolipoprotein D and pepsinogen C) in ductal carcinoma in situ of the breast. Histopathology 2007; 50:866-74. [PMID: 17543076 DOI: 10.1111/j.1365-2559.2007.02687.x] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/01/2022]
Abstract
AIMS To evaluate the expression of androgen receptors (AR) and two androgen-induced proteins [apolipoprotein D (ApoD) and pepsinogen C (PepC)] in ductal carcinoma in situ (DCIS) of the breast. METHODS AND RESULTS AR, ApoD and PepC expression was examined in 28 cases of pure DCIS and in 31 cases of DCIS adjacent to invasive carcinoma of the breast using immunohistochemical methods and then correlated with the architectural subtype, the degree of differentiation and the ostrogen receptor (ER)/progesterone receptor (PgR)/HER-2 status. We found no significant differences between pure DCIS and DCIS adjacent to invasive breast cancer regarding the percentage of positive cases for ApoD (64.3% versus 54.8%), PepC (42.9% versus 48.4%), ER (64.3% versus 58.1%), PgR (60.7% versus 58.1%) and HER-2 (39.3% versus 67.7%). However, there was a significantly higher percentage of AR+ DCIS among those adjacent to invasive carcinomas of the breast than among pure DCIS lesions (93.5% versus 60.9%) (P = 0.009). AR expression did not correlate with architectural subtype, degree of differentiation, or ER/PgR/HER-2/ApoD/PepC status, in cases of pure DCIS, nor in DCIS adjacent to invasive carcinoma of the breast. CONCLUSIONS AR expression may represent an independent predictive factor in DCIS of the breast.
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MESH Headings
- Apolipoproteins D/metabolism
- Biomarkers, Tumor/metabolism
- Breast Neoplasms/metabolism
- Breast Neoplasms/pathology
- Carcinoma, Ductal, Breast/metabolism
- Carcinoma, Ductal, Breast/pathology
- Carcinoma, Intraductal, Noninfiltrating/metabolism
- Carcinoma, Intraductal, Noninfiltrating/pathology
- Female
- Glycoproteins/metabolism
- Humans
- Immunohistochemistry
- Membrane Transport Proteins/metabolism
- Middle Aged
- Neoplasm Proteins/metabolism
- Pepsinogen C/metabolism
- Receptor, ErbB-2/metabolism
- Receptors, Androgen/metabolism
- Receptors, Estrogen/metabolism
- Receptors, Progesterone/metabolism
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Affiliation(s)
- L O Gonzalez
- Instituto Universitario de Oncologí,a del Principado de Asturias, Hospital de Jove, Gijón, Spain
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12
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Vizoso FJ, Rodriguez M, Altadill A, González-Diéguez ML, Linares A, González LO, Junquera S, Fresno-Forcelledo F, Corte MD, Rodrigo L. Liver expression of steroid hormones and Apolipoprotein D receptors in hepatocellular carcinoma. World J Gastroenterol 2007; 13:3221-3227. [PMID: 17589901 PMCID: PMC4436608 DOI: 10.3748/wjg.v13.i23.3221] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/21/2007] [Revised: 03/05/2007] [Accepted: 03/15/2007] [Indexed: 02/06/2023] Open
Abstract
AIM To evaluate the tissular expression of Androgen (A), Estrogen (E) and Progesterone (Pg) receptors, and Apolipoprotein D (ApoD), in liver tumors from resected hepatocellular carcinoma (HCC) cases in order to assess their possible relationship to prognosis. METHODS We performed an immunohistochemical study using tissue microarrays (containing more than 260 cancer specimens, from 31 HCC patients and controls) to determine the presence of specific antibodies against AR, ER, PgR and ApoD, correlating their findings with several clinico-pathological and biological variables. The staining results were categorized using a semi-quantitative score based on their intensity, and the percentage of immunostained cells was measured. RESULTS A total of 21 liver tumors (67.7%) were positive for AR; 16 (51.6%) for ER; 26 (83.9%) for PgR and 12 (38.7%) stained for ApoD. We have found a wide variability in the immunostaining score values for each protein, with a median (range) of 11.5 (11.5-229.5) for AR; 11.1 (8.5-65) for ER; 14.2 (4-61) for PgR; and 37.7 (13.8-81.1) for ApoD. A history of heavy ethanol consumption, correlated positively with AR and PgR and negatively with ER status. HCV chronic infection also correlated positively with AR and PgR status. However, the presence of ApoD immunostaining did not correlate with any of these variables. Tumors with a positive immuno-staining for PgR showed a better prognosis. CONCLUSION Our results indicate a moderate clinical value of the steroid receptor status in HCC, emphasizing the need to perform further studies in order to evaluate the possible role of new hormonal-based therapies.
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Affiliation(s)
- F J Vizoso
- Unidad de Investigación, Fundación Hospital de Jove, Gijón, and Instituto Universitario de Oncología del Principado de Asturias, Oviedo, Spain
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13
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Russell ST, Tisdale MJ. Effect of eicosapentaenoic acid (EPA) on expression of a lipid mobilizing factor in adipose tissue in cancer cachexia. Prostaglandins Leukot Essent Fatty Acids 2005; 72:409-14. [PMID: 15899583 DOI: 10.1016/j.plefa.2005.03.002] [Citation(s) in RCA: 36] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/04/2005] [Revised: 03/16/2005] [Accepted: 03/24/2005] [Indexed: 11/25/2022]
Abstract
Adipose tissue of mice bearing a cachexia-inducing murine tumour (MAC16) shows increased expression of zinc-alpha(2)-glycoprotein (ZAG), a lipolytic factor thought to be responsible for the increased lipolysis. The anti-cachectic agent eicosapentaenoic acid (EPA) (0.5 g/kg) attenuated the loss of body weight in mice bearing the MAC16 tumour, and this was accompanied by downregulation of ZAG expression in both white and brown adipose tissue, as determined by Western blotting. Glucocorticoids may be responsible for the increased ZAG expression in adipose tissue. Dexamethasone (1.68 microM) stimulated lipolysis in 3T3-L1 adipocytes, and this effect was attenuated by EPA (50 microM). In addition the lipolytic action of dexamethasone was attenuated by anti-ZAG antibody, suggesting that the induction of lipolysis was mediated through an increase in ZAG expression. This was confirmed by Western blotting, which showed that dexamethasone (1.68 microM) induced a two-fold increase in ZAG expression in both cells and media, and that this was attenuated by EPA (50 microM). These results suggest that EPA may preserve adipose tissue in cachectic mice by downregulation of ZAG expression through interference with glucocorticoid signalling.
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Affiliation(s)
- S T Russell
- Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, UK
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Mrusek S, Classen-Linke I, Vloet A, Beier HM, Krusche CA. Estradiol and medroxyprogesterone acetate regulated genes in T47D breast cancer cells. Mol Cell Endocrinol 2005; 235:39-50. [PMID: 15866426 DOI: 10.1016/j.mce.2005.01.009] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/23/2004] [Revised: 12/21/2004] [Accepted: 01/17/2005] [Indexed: 11/26/2022]
Abstract
Many mammary tumors express estrogen receptors (ER) and progesterone receptors (PR), and there is increasing evidence that progestins influence gene expression of breast tumor cells. To analyse the impact of progestins on breast cancer cells, we compared (a) the expression of two cytokines, involved in tumor progression, and searched (b) for differentially regulated genes by a microarray, containing 2400 genes, on T47D breast cancer cells cultured for 6 days with 17beta-estradiol (E2) or E2+medroxyprogesterone acetate (E2+MPA). Lower amounts of PDGF and TNFalpha were found in culture supernatants of E2+MPA treated T47D cells. MPA addition induced a 2.8-3.5-fold increase of the mRNA expression of (a) tristetraprolin, which is involved in the posttranscriptional regulation of cytokine biosynthesis, and (b) zinc-alpha2-glycoprotein and Na, K-ATPase alpha1-subunit, which both resemble differentiation markers of breast epithelium. In contrast, the mRNA expression of lipocalin 2, which promotes matrixmetalloproteinase-9 activity, was decreased five-fold in E2+MPA treated cells. Our data show that the expression of genes from various functional gene families is regulated differentially by E2 and E2+MPA treatment in T47D cells. This suggests that exogenous progestins applied for therapy and endogenous changes of the progesterone levels during the menstrual cycle both influence breast cancer pathophysiology.
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Affiliation(s)
- S Mrusek
- Department of Anatomy and Reproductive Biology, RWTH Aachen University, Wendlingweg 2, 52074 Aachen, Germany
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15
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Russell ST, Tisdale MJ. The role of glucocorticoids in the induction of zinc-alpha2-glycoprotein expression in adipose tissue in cancer cachexia. Br J Cancer 2005; 92:876-81. [PMID: 15714206 PMCID: PMC2361908 DOI: 10.1038/sj.bjc.6602404] [Citation(s) in RCA: 78] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022] Open
Abstract
Loss of adipose tissue in cancer cachexia in mice bearing the MAC16 tumour arises from an increased lipid mobilisation through increased expression of zinc-α2-glycoprotein (ZAG) in white (WAT) and brown (BAT) adipose tissue. Glucocorticoids have been suggested to increase ZAG expression, and this study examines their role in cachexia and the mechanisms involved. In mice bearing the MAC16 tumour, serum cortisol concentrations increased in parallel with weight loss, and the glucocorticoid receptor antagonist RU38486 (25 mg kg−1) attenuated both the loss of body weight and ZAG expression in WAT. Dexamethasone (66 μg kg−1) administration to normal mice produced a six-fold increase in ZAG expression in both WAT and BAT, which was also attenuated by RU38486. In vitro studies using 3T3-L1 adipocytes showed dexamethasone (1.68 μM) to stimulate lipolysis and increase ZAG expression, and both were attenuated by RU38486 (10 μM), anti-ZAG antibody (1 μgml−1), and the β3-adrenoreceptor (β3-AR) antagonist SR59230A (10 μM). Zinc-α2-glycoprotein also increased its own expression and this was attenuated by SR59230A, suggesting that it was mediated through the β3-AR. This suggests that glucocorticoids stimulate lipolysis through an increase in ZAG expression, and that they are responsible for the increase in ZAG expression seen in adipose tissue of cachectic mice.
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Affiliation(s)
- S T Russell
- Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, UK
| | - M J Tisdale
- Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, UK
- Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, UK. E-mail:
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Bao Y, Bing C, Hunter L, Jenkins JR, Wabitsch M, Trayhurn P. Zinc-alpha2-glycoprotein, a lipid mobilizing factor, is expressed and secreted by human (SGBS) adipocytes. FEBS Lett 2005; 579:41-7. [PMID: 15620688 DOI: 10.1016/j.febslet.2004.11.042] [Citation(s) in RCA: 142] [Impact Index Per Article: 7.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2004] [Revised: 11/09/2004] [Accepted: 11/13/2004] [Indexed: 11/24/2022]
Abstract
Zinc-alpha2-glycoprotein (ZAG), a lipid mobilizing factor, is expressed in mouse adipose tissue and is markedly upregulated in mice with cancer cachexia. We have explored whether ZAG is expressed and secreted by human adipocytes, using SGBS cells, and examined the regulation of ZAG expression. ZAG mRNA was detected by RT-PCR in mature human adipocytes and in SGBS cells post-, but not pre-, differentiation to adipocytes. Relative ZAG mRNA levels increased rapidly after differentiation of SGBS cells, peaking at day 8 post-induction. ZAG protein was evident in differentiated adipocytes (by day 3) and also detected in the culture medium (by day 6) post-induction. The PPARgamma agonist rosiglitazone induced a 3-fold increase in ZAG mRNA level, while TNF-alpha led to a 4-fold decrease. Human adipocytes express and secrete ZAG, with ZAG expression being regulated particularly through TNF-alpha and the PPARgamma nuclear receptor. ZAG is a novel adipokine, which may be involved in the local regulation of adipose tissue function.
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Affiliation(s)
- Yi Bao
- Neuroendocrine & Obesity Biology Unit, School of Clinical Sciences, University Clinical Departments, University of Liverpool, Liverpool L69 3GA, UK
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17
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Bing C, Bao Y, Jenkins J, Sanders P, Manieri M, Cinti S, Tisdale MJ, Trayhurn P. Zinc-alpha2-glycoprotein, a lipid mobilizing factor, is expressed in adipocytes and is up-regulated in mice with cancer cachexia. Proc Natl Acad Sci U S A 2004; 101:2500-2505. [PMID: 14983038 PMCID: PMC356979 DOI: 10.1073/pnas.0308647100] [Citation(s) in RCA: 228] [Impact Index Per Article: 10.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/28/2003] [Indexed: 01/23/2023] Open
Abstract
Zinc-alpha2-glycoprotein (ZAG), a 43-kDa protein, is overexpressed in certain human malignant tumors and acts as a lipid-mobilizing factor to stimulate lipolysis in adipocytes leading to cachexia in mice implanted with ZAG-producing tumors. Because white adipose tissue (WAT) is an endocrine organ secreting a wide range of protein factors, including those involved in lipid metabolism, we have investigated whether ZAG is produced locally by adipocytes. ZAG mRNA was detected by RT-PCR in the mouse WAT depots examined (epididymal, perirenal, s.c., and mammary gland) and in interscapular brown fat. In WAT, ZAG gene expression was evident in mature adipocytes and in stromal-vascular cells. Using a ZAG Ab, ZAG protein was located in WAT by Western blotting and immunohistochemistry. Mice bearing the MAC16-tumor displayed substantial losses of body weight and fat mass, which was accompanied by major increases in ZAG mRNA and protein levels in WAT and brown fat. ZAG mRNA was detected in 3T3-L1 cells, before and after the induction of differentiation, with the level increasing progressively after differentiation with a peak at days 8-10. Both dexamethasone and a beta3 agonist, BRL 37344, increased ZAG mRNA levels in 3T3-L1 adipocytes. ZAG gene expression and protein were also detected in human adipose tissue (visceral and s.c.). It is suggested that ZAG is a new adipose tissue protein factor, which may be involved in the modulation of lipolysis in adipocytes. Overexpression in WAT of tumor-bearing mice suggests a local role for adipocyte-derived ZAG in the substantial reduction of adiposity of cancer cachexia.
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Affiliation(s)
- Chen Bing
- Neuroendocrine and Obesity Biology Unit, Department of Medicine, University Clinical Departments, University of Liverpool, Liverpool L69 3GA, UK
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18
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Miranda E, Vizoso F, Martín A, Quintela I, Corte MD, Seguí ME, Ordiz I, Merino AM. Apolipoprotein D expression in cutaneous malignant melanoma. J Surg Oncol 2003; 83:99-105. [PMID: 12772203 DOI: 10.1002/jso.10245] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
BACKGROUND AND OBJECTIVES Apolipoprotein D (Apo D) is a protein component of the human plasma lipid transport system, and an androgen-regulated protein in both breast and prostate cancer cell lines. Our goal was to evaluate the expression of Apo D in malignant cutaneous melanomas, as well as to assess its possible relationship to clinical and pathological parameters. METHODS Apo D expression was analyzed in 32 paraffin-embedded tissues from patients with invasive cutaneous malignant melanomas, in 8 samples from in situ melanoma, and in 10 samples from 10 benign lesions (4 dermal melanocytic nevi, 4 compound melanocytic nevi, and 2 dysplastic melanocytic nevi), using immunohistochemical techniques. RESULTS The benign lesions were consistently negative for Apo D, whereas 3 of the 8 "in situ" melanomas (37.5%) and 12 of the 32 invasive melanomas (37.5%) showed positive immunostaining for Apo D. The percentage of Apo D-positive tumors was significantly higher in nodular than in superficial spreading melanomas (P = 0.011) and in melanomas with vertical growth phase than in melanomas with radial growth phase (P = 0.02). In addition, the percentage of Apo D-positive tumors was positively and significantly correlated with Clark's level of invasion (P = 0.046). CONCLUSIONS Apo D may be a new prognostic factor of unfavorable evolution in cutaneous malignant melanoma.
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Affiliation(s)
- Eva Miranda
- Department of Pathology. Hospital de Cabueñes, Gijón, Spain
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19
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Abstract
Zinc alpha-2-glycoprotein is secreted by a variety of normal and malignant epithelial cells and overexpression by tumors has been implicated in cancer cachexia. To investigate biologic properties of zinc alpha-2-glycoprotein further, stable transfectants of recombinant human zinc alpha-2-glycoprotein were created in the B16F10 murine melanoma cell line. Both B16-recombinant human zinc alpha-2-glycoprotein clones with strong expression of zinc alpha-2-glycoprotein and vector-transfected B16 cells treated with exogenous zinc alpha-2-glycoprotein had decreased melanin production in vitro. Furthermore, B16-recombinant human zinc alpha-2-glycoprotein clones formed amelanotic tumors in vivo, despite their melanin production in vitro. Although no qualitative differences in tyrosinase mRNA expression could be detected by reverse transcription-polymerase chain reaction, B16-recombinant human zinc alpha-2-glycoprotein tumors had decreased levels of tyrosinase protein and minimal tyrosinase activity. Purified zinc alpha-2-glycoprotein also decreased tyrosinase activity in vector-transfected B16 tumor sections in vitro. Taken together, these studies demonstrate that zinc alpha-2-glycoprotein inhibits melanin production by B16 melanoma cells via post-transcriptional effects on tyrosinase protein. As zinc alpha-2-glycoprotein decreases melanin synthesis more strongly in vivo than in vitro, however, it is likely that zinc alpha-2-glycoprotein affects melanin synthesis through indirect mechanisms as well. Zinc alpha-2-glycoprotein also inhibits melanin production by melan-A primary melanocytes in vitro. As zinc alpha-2-glycoprotein is normally produced by epidermal keratinocytes, these studies raise the possibility that epidermal-derived zinc alpha-2-glycoprotein may play a part in normal regulation of melanin production in vivo, in addition to its previously described role in cancer cachexia.
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Affiliation(s)
- Laura P Hale
- Department of Pathology and the Comprehensive Cancer Center, Duke University Medical Center, Durham, North Carolina 27710, USA.
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20
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Rojo JV, González LO, Lamelas ML, Merino A, Vizoso F. Apolipoprotein D expression in endometrial carcinomas. Acta Obstet Gynecol Scand 2001; 80:158-61. [PMID: 11167212 DOI: 10.1034/j.1600-0412.2001.080002158.x] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/25/2022]
Abstract
BACKGROUND Apolipoprotein D is a protein component of the human plasma lipid transport system but is also associated with a more favorable prognosis in breast cancer and ovarian cancer women. This study was undertaken to examine the tumoral expression of apolipoprotein D in endometrial cancer and to analyze the possible correlation with tumor and patients characteristics as well as androgen receptors and its prognostic significance. METHODS Immunohistochemical evaluation was used to examine apolipoprotein D expression in paraffin blocks from 58 endometrial carcinomas. RESULTS A total of twenty (34%) tumors stained positively. Staining was localized in tumor cells. No significant correlation was found between apo D expression and patients or tumor characteristics and androgen receptor status. In addition, apolipoprotein D expression was not associated with patient prognosis. CONCLUSIONS Apolipoprotein D is expressed by a significant percentage of endometrial carcinomas without apparent association with other clinicopathologic parameters or with outcome of patients.
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Affiliation(s)
- J V Rojo
- Department of Gynecology, Hospital de Jove, Gijón, Asturias, Spain
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21
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Vázquez J, González L, Merino A, Vizoso F. Expression and clinical significance of apolipoprotein D in epithelial ovarian carcinomas. Gynecol Oncol 2000; 76:340-7. [PMID: 10684708 DOI: 10.1006/gyno.1999.5678] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
OBJECTIVE Apolipoprotein D is a protein component of the human plasma lipid transport system but is also associated with a more favorable prognosis in women with breast cancer. This retrospective study was undertaken to examine the tumoral expression of apolipoprotein D in epithelial ovarian cancer and to analyze the possible correlation with tumor and patient characteristics as well as androgen receptors and their prognostic significance. METHODS Immunohistochemical evaluation was used to examine apolipoprotein D expression in paraffin blocks from 68 epithelial ovarian carcinomas. RESULTS A total of 18 (26.4%) tumors stained positively. No significant correlation was found between apolipoprotein D expression and patient or tumor characteristics and androgen receptor status. However, apolipoprotein D expression was significantly associated with prognosis in patients with residual tumor greater than 1 cm. Thus, patients with apolipoprotein-D-negative tumors had a poorer overall survival than those with apolipoprotein-D-positive tumors (P = 0.039). In addition, multivariate analysis demonstrated that apolipoprotein D expression was an independent prognostic factor with initial tumor size in this group of patients (P = 0.005). CONCLUSIONS Our results led us to consider the existence of apolipoprotein D expression by a significative percentage of ovarian carcinomas, and this protein expression might be of clinical usefulness for identifying lesions with different evolution.
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Affiliation(s)
- J Vázquez
- Department of Gynecology, Hospital de Jove, Gijón, Spain
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22
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Walker MG, Volkmuth W, Sprinzak E, Hodgson D, Klingler T. Prediction of gene function by genome-scale expression analysis: prostate cancer-associated genes. Genome Res 1999; 9:1198-203. [PMID: 10613842 PMCID: PMC310991 DOI: 10.1101/gr.9.12.1198] [Citation(s) in RCA: 130] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
We wish to identify genes associated with disease. To do so, we look for novel genes whose expression patterns mimic those of known disease-associated genes, using a method we call Guilt-by-Association (GBA), on the basis of a combinatoric measure of association. Using GBA, we have examined the expression of 40,000 human genes in 522 cDNA libraries, and have discovered several hundred previously unidentified genes associated with cancer, inflammation, steroid-synthesis, insulin-synthesis, neurotransmitter processing, matrix remodeling, and other disease processes. The majority of the genes thus discovered show no sequence similarity to known genes, and thus could not have been identified by homology searches. We present here an example of the discovery of eight genes associated with prostate cancer. Of the 40,000 most-abundant human genes, these 8 are the most closely linked to the known diagnostic genes, and thus are prime targets for pharmaceutical research.
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Affiliation(s)
- M G Walker
- Incyte Pharmaceuticals, Palo Alto, California 94304, USA.
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23
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Lei G, Brysk H, Arany I, Tyring SK, Srinivasan G, Brysk MM. Characterization of zinc-?2-glycoprotein as a cell adhesion molecule that inhibits the proliferation of an oral tumor cell line. J Cell Biochem 1999. [DOI: 10.1002/(sici)1097-4644(19991001)75:1<160::aid-jcb16>3.0.co;2-b] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
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Sánchez LM, Chirino AJ, Bjorkman PJ. Crystal structure of human ZAG, a fat-depleting factor related to MHC molecules. Science 1999; 283:1914-9. [PMID: 10206894 DOI: 10.1126/science.283.5409.1914] [Citation(s) in RCA: 110] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
Zn-alpha2-glycoprotein (ZAG) is a soluble protein that is present in serum and other body fluids. ZAG stimulates lipid degradation in adipocytes and causes the extensive fat losses associated with some advanced cancers. The 2.8 angstrom crystal structure of ZAG resembles a class I major histocompatibility complex (MHC) heavy chain, but ZAG does not bind the class I light chain beta2-microglobulin. The ZAG structure includes a large groove analogous to class I MHC peptide binding grooves. Instead of a peptide, the ZAG groove contains a nonpeptidic compound that may be implicated in lipid catabolism under normal or pathological conditions.
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Affiliation(s)
- L M Sánchez
- Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA
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25
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López-Otín C, Diamandis EP. Breast and prostate cancer: an analysis of common epidemiological, genetic, and biochemical features. Endocr Rev 1998; 19:365-96. [PMID: 9715372 DOI: 10.1210/edrv.19.4.0337] [Citation(s) in RCA: 45] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Affiliation(s)
- C López-Otín
- Departamento de Bioquímica, Facultad de Medicina, Universidad de Oviedo, Spain
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de Bolòs C, Gumà M, Barranco C, Garrido M, Kim YS, Real FX. MUC6 expression in breast tissues and cultured cells: abnormal expression in tumors and regulation by steroid hormones. Int J Cancer 1998; 77:193-9. [PMID: 9650551 DOI: 10.1002/(sici)1097-0215(19980717)77:2<193::aid-ijc4>3.0.co;2-l] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
Neoplastic transformation of epithelial cells is commonly associated with alterations in the expression of mucin genes. The mechanisms involved in this process are largely unknown. MUC6, isolated from a stomach cDNA library, is mainly expressed in stomach antral glands, as detected by using in situ hybridization and immunohistochemistry. We examined MUC6 expression in normal and pathological breast tissues using immunohistochemistry with MUC6-specific antibodies and in cultured breast cancer cells using immunocytochemistry and Northern blotting. MUC6 was generally not detected in normal breast (1/11) but was detected in fibrocystic disease without atypia (7/17, 41%), in atypical fibrocystic disease (11/11, 100%) and in carcinoma (57/60, 95%). To study the mechanisms involved in mucin gene up-regulation in breast cancer, we examined baseline, growth-related and steroid-induced levels of MUC1, MUC3 and MUC6 in 4 breast cancer cell lines, 2 of which express estrogen receptors. MUC6 levels were up-regulated at post-confluence in 2/4 cell lines, whereas no changes were detected for the other mucin genes examined. MUC6 and MUC3 were constitutively expressed, and steroid-induced, in BT-474 and MCF-7 cells, respectively. As a control, pS2 was induced in both cell lines. Our results indicate that (1) MUC6 is overexpressed in breast cancer and in benign breast disease, (2) in vitro, MUC6 and MUC3 are up-regulated by steroids and (3) abnormal expression of MUC6 in breast cancers may, in part, be explained by hormonal changes associated with tumor development.
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Affiliation(s)
- C de Bolòs
- Unitat de Biologia Cel.lular i Molecular, Institut Municipal d'Investigació Mèdica, Barcelona, Spain
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Karpenko V, Kaupová M, Kodícek M. The conformation and stability of human Zn-alpha 2-glycoprotein in aqueous and methanolic solutions. Biophys Chem 1997; 69:209-17. [PMID: 9474755 DOI: 10.1016/s0301-4622(97)00093-8] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
The stability of Zn-alpha 2-glycoprotein has been studied using UV-spectroscopy and circular dichroism with respect to the influence of temperature, pH, and solvent composition. It has been found that: (1) this protein contains a relatively high proportion of beta-sheet (60%) and a very low amount of other periodic structures as estimated from circular dichroic spectra; (2) at pH 7.4, the circular dichroic spectra change reversibly in the temperature range between 25 and 85 degrees C; small disturbances were observed at 265 nm; (3) with the assumption of the two-state process, the temperature of cooperative denaturation was Tm = 66 degrees C, van't Hoff's enthalpy of this process was 27 kJ mol-1; (4) up to pH 9.5 the dichroic spectrum appeared the same as at pH 7.4; (5) in the presence of methanol (vol. fraction 50%), no isodichroic points on the circular dichroic spectra were found during temperature denaturation; after cooling from 85 degrees C, the alpha-helix content was higher than in the native protein; (6) in the molecule of Zn-alpha 2-glycoprotein, 14 out of 18 tyrosines can dissociate with the instrinsic pK = 11.2; and (7) the temperature perturbation difference spectra yielded nonlinear delta A vs. T curves with temperature transition corresponding to the values found in the circular dichroic spectra; the numbers of chromophores exposed to the solvent as determined by the temperature difference spectra were: 4 tyrosines, 1 tryptophan, and 1 phenylalanine. In several aspects, a parallel has been found between Zn-alpha 2-glycoprotein and orosomucoid (acid alpha 1-glycoprotein), another plasma glycoprotein.
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Affiliation(s)
- V Karpenko
- Charles University, Faculty of Natural Sciences, Department of Physical and Macromolecular Chemistry, Prague, Czech Republic.
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Juricskay S, Szabó I, Kett K. Urinary steroids at time of surgery in postmenopausal women with breast cancer. Breast Cancer Res Treat 1997; 44:83-9. [PMID: 9164681 DOI: 10.1023/a:1005749032226] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/04/2023]
Abstract
Urinary steroid metabolites were measured by capillary gas chromatography in 22 postmenopausal women with operable breast cancer on day before the tumour excision and in 20 hospitalised control who were before an operation from other cause than cancer. Serum dehydroepiandrosterone-sulphat (DHEAS) and testosterone (T) level were measured by radioimmunassay in the same groups and same time. There was no significant difference in the level of urinary androgen metabolites. Pregnanediol level was significantly lower (P < 0.05) in cancer patients. In the 5 patients with positive axillary nodes the tetrahydrocortisol and alpha-cortolone levels were significantly (P < 0.05) higher than in node negative ones. There was no significant differences in the serum DHEAS and T levels. These results indicate that metabolic changes are existing in postmenopausal patients which may be a cause or a consequence of the disease.
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Affiliation(s)
- S Juricskay
- Central Research Laboratory, Medical University of Pécs, Hungary.
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Sánchez LM, López-Otín C, Bjorkman PJ. Biochemical characterization and crystalization of human Zn-alpha2-glycoprotein, a soluble class I major histocompatibility complex homolog. Proc Natl Acad Sci U S A 1997; 94:4626-30. [PMID: 9114041 PMCID: PMC20774 DOI: 10.1073/pnas.94.9.4626] [Citation(s) in RCA: 47] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/1996] [Accepted: 02/21/1997] [Indexed: 02/04/2023] Open
Abstract
Zn-alpha2-glycoprotein (ZAG) is a 41-kDa soluble protein that is present in most bodily fluids. In addition, ZAG accumulates in fluids from breast cysts and in 40% of breast carcinomas, which suggests that ZAG plays a role in the development of breast diseases. However, the function of ZAG under physiological and cancerous conditions remains unknown. Because ZAG shares 30-40% sequence identity with the heavy chains of class I major histocompatibility complex (MHC) proteins, we compared the biochemical properties of ZAG with those of classical class I MHC molecules. We purified human ZAG from breast cyst fluid and serum and produced a panel of anti-ZAG monoclonal antibodies. Binding assays and acid elution experiments revealed that, in contrast to class I MHC proteins, ZAG does not bind peptides or the class I light chain, beta2-microglobulin (beta2m). Nevertheless, CD studies indicated that ZAG is thermally stable in the absence of bound peptide or associated beta2m, as opposed to class I MHC molecules, which require the presence of both beta2m and peptides for stability. These data indicate that the function of ZAG has diverged from the peptide presentation and T-cell interaction functions of class I molecules. To gain insight into the function of ZAG and to compare the three-dimensional structures of ZAG and class I MHC molecules, we produced ZAG crystals that diffract beyond 2.7 A and have initiated an x-ray structure determination.
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Affiliation(s)
- L M Sánchez
- Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA
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Balbín M, López-Otín C. Hormonal regulation of the human pepsinogen C gene in breast cancer cells. Identification of a cis-acting element mediating its induction by androgens, glucocorticoids, and progesterone. J Biol Chem 1996; 271:15175-81. [PMID: 8663058 DOI: 10.1074/jbc.271.25.15175] [Citation(s) in RCA: 37] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/01/2023] Open
Abstract
Pepsinogen C is an aspartic proteinase mainly involved in the digestion of proteins in the stomach, which is also synthesized by certain human breast tumors. To examine the possibility that extragastric production of this proteolytic enzyme could be mediated by hormonal factors, we have analyzed pepsinogen C gene expression in human breast cancer cells subjected to different hormonal treatments. Northern blot analyses revealed the expression of pepsinogen C gene by T-47D breast cancer cells after induction with dihydrotestosterone, dexamethasone, and progesterone but not with estradiol, retinoic acid, or ethanol. Reverse transcription-polymerase chain reaction analysis in a series of breast cancer cell lines confirmed the amplification of pepsinogen C mRNA after induction with dihydrotestosterone, in those cells expressing the androgen receptor mRNA. The promoter region of the pepsinogen C gene was functionally characterized by transient expression of a vector containing the promoter region cloned in front of the chloramphenicol acetyltransferase (CAT) reporter gene. CAT activity in T-47D cells was stimulated in the presence of dihydrotestosterone, dexamethasone, and progesterone but not by estradiol. By further deletion mapping of the pepsinogen C promoter, a minimal region (AGAACTattTGTTCC) was identified as being responsible for glucocorticoid-, androgen-, and progesterone-regulated gene expression.
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Affiliation(s)
- M Balbín
- Departamento de Biología Funcional, Universidad de Oviedo, 33006 Oviedo, Spain
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van der Pompe G, Antoni MH, Heijnen CJ. Elevated basal cortisol levels and attenuated ACTH and cortisol responses to a behavioral challenge in women with metastatic breast cancer. Psychoneuroendocrinology 1996; 21:361-74. [PMID: 8844875 DOI: 10.1016/0306-4530(96)00009-1] [Citation(s) in RCA: 75] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/02/2023]
Abstract
Hormones of the hypothalamic-pituitary-adrenal system were studied in 31 patients with early stage breast cancer and patients with metastatic breast cancer. Both groups received tamoxifen as first-line treatment. As a control group 15 age-matched healthy women participated in the study. The results showed that breast cancer patients had significant elevations in basal cortisol levels compared to controls. Metastatic breast cancer patients had higher cortisol levels than early stage breast cancer patients. No significant differences between breast cancer patients and controls were found in basal plasma ACTH and prolactin levels. These data provide evidence that breast cancer is associated with a hyperactive adrenal gland, which may be due to the physiological stress associated with the presence of (micro)metastases or tumor cells in the circulation, in combination with administration of tamoxifen. In response to a behavioral challenge increases were observed in plasma ACTH and prolactin. Metastatic breast cancer patients had a faster prolactin response to acute stress than healthy women. However, metastatic breast cancer patients showed a blunted ACTH response compared to healthy women. Stress-induced ACTH responses and basal cortisol levels were negatively correlated in the metastatic group only. Thus, the blunted ACTH response to the behavioral challenge might be related to hypercortisolemia suggesting that the pituitary corticotroph cell in metastatic cancer is appropriately restrained possibly by the negative feedback effects of chronic cortisol elevations. Interestingly, the behavioral challenge induced decreases in cortisol levels in all three groups. However, metastatic breast cancer patients had a faster cortisol decline compared to healthy women. We hypothesize that this is caused by increased metabolic clearance of cortisol due to increased utilization of metabolic substrates often observed in the presence of a tumor.
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Affiliation(s)
- G van der Pompe
- Helen Dowling Institute for Biopsychosocial Medicine, Rotterdam, The Netherlands.
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Apolipoprotein D gene induction by retinoic acid is concomitant with growth arrest and cell differentiation in human breast cancer cells. J Biol Chem 1994. [DOI: 10.1016/s0021-9258(18)47100-1] [Citation(s) in RCA: 52] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022] Open
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