Nowadays, diet and breast cancer are studied at different levels, particularly in tumor prevention and progression. Thus, the molecular mechanisms leading to better knowledge are deciphered with a higher precision. Among the molecules implicated in a preventive and anti-progressive way, n-3 long chain polyunsaturated fatty acids (n-3 LC-PUFAs) are good candidates. These molecules, like docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids, are generally found in marine material, such as fat fishes or microalgae. EPA and DHA act as anti-proliferative, anti-invasive, and anti-angiogenic molecules in breast cancer cell lines, as well as in in vivo studies. A better characterization of the cellular and molecular pathways involving the action of these fatty acids is essential to have a realistic image of the therapeutic avenues envisaged behind their use. This need is reinforced by the increase in the number of clinical trials involving more and more n-3 LC-PUFAs, and this, in various pathologies ranging from obesity to a multitude of cancers. The objective of this review is, therefore, to highlight the new elements showing the preventive and beneficial effects of n-3 LC-PUFAs against the development and progression of breast cancer.

15-lipoxygenase is one of the key enzymes for the metabolism of unsaturated fatty acids that its manipulation has been proposed recently as a new molecular target for regulating cancer cell growth. Aberrant expression of 15-lipoxygenase enzyme seems to play an indicative role in the pathology of different cancer types, tumor progression, metastasis, or apoptosis. Based on the fact that breast cancer is one of the most common cancers that imposes a burden of mortality in women also, on the other hand, evidence in experimental models and human studies indicate the emerging role of the 15-lipoxygenase pathway in breast cancer pathogenesis, we present a review of recent findings related to the role of 15- lipoxygenase enzyme and metabolites in breast cancer growth, apoptosis, metastasis, and invasion as well as their local and circulating expression pattern in patients with breast cancer. Our review supports the emerging role of 15- lipoxygenase in molecular and cellular processes regulating breast tumor cell fate with both positive and negative effects.

• 15-lipoxygenase-1
• 15-lipoxygenase-2
• Apoptosis
• Breast cancer
• Cell growth
• Metastasis

• Breast cancer
• Crosstalk
• Ferroptosis
• Ferroptotic regulators
• Historical perspective
• Regulated cell death

• 81972453 National Natural Science Foundation of China
• 81602471 National Natural Science Foundation of China
• 81972597 National Natural Science Foundation of China
• 81672729 National Natural Science Foundation of China
• LY19H160055 Natural Science Foundation of Zhejiang Province
• LY19H160059 Natural Science Foundation of Zhejiang Province
• Zheng Shu Medical Elite Scholarship Fund

n-3 long chain Polyunsaturated Fatty Acids (n-3 LCPUFA) have been shown to improve the efficacy of conventional chemotherapies used for breast cancer treatment. In addition to their reported ability to increase the chemosensitivity of cancer cells, we hypothesized that n-3 LCPUFA could induce a remodeling of the vascular network in mammary tumors. A contrast-enhanced ultrasound method was used to monitor the vascular architecture during docetaxel treatment of mammary tumors in rats fed either a control or an n-3 LCPUFA-enriched diet (docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA)). The vascular network was remodeled in favor of smaller vessels (microvascularization), which represented 54% of the vasculature in n-3 LCPUFA tumors but only 26% in control tumors after 2 weeks of chemotherapy. Importantly, vascularization changes occurred both before and during docetaxel treatment. The density of smaller vessels quantified before chemotherapy was correlated with improved tumor size reduction by docetaxel treatment. Furthermore, transcript levels of the angiogenesis-specific genes epiregulin and amphiregulin were reduced by ~4.5- and twofold in tumors obtained from rats fed an n-3 LCPUFA-enriched diet compared to those of rats fed a control diet, respectively. Their expression levels were negatively correlated with tumor regression after chemotherapy. Taken together, this preclinical data strengthen the potential usefulness of n-3 LCPUFA as a complementary clinical strategy to improve drug efficiency via remodeling of the tumor vasculature.

• amphiregulin
• docetaxel
• epiregulin
• mammary tumors
• n-3 PUFA
• vascularization

• Animals
• Diet
• Docetaxel/pharmacology
• Docosahexaenoic Acids/pharmacology
• Eicosapentaenoic Acid/pharmacology
• Fatty Acids/pharmacology
• Fatty Acids, Omega-3/pharmacology
• Female
• Mammary Neoplasms, Animal/drug therapy
• Mammary Neoplasms, Animal/pathology
• Neovascularization, Pathologic/drug therapy
• Neovascularization, Pathologic/metabolism
• Rats
• Rats, Sprague-Dawley

• Aged
• Carcinoma, Squamous Cell/metabolism
• Carcinoma, Squamous Cell/pathology
• Chromatography, Gas
• Fatty Acids/metabolism
• Female
• Humans
• Male
• Middle Aged
• Mouth Neoplasms/metabolism
• Mouth Neoplasms/pathology
• Tumor Microenvironment

Vascular Smooth Muscle cells (VSMC) enact crucial roles in early vasculogenesis and sustenance of vascular integrity. However, aberrant proliferation of VSMC followed by migration into the blood vessel wall leads to the formation of vascular lesions accounting for the degeneration and remodelling of vascular basement membrane. In diabetes, hyperglycaemia accelerates VSMC proliferation and contributes to the initiation and progression of atherosclerotic lesions. Recently, acute glucose fluctuations have been implicated in the abnormal VSMC proliferation and complications of diabetic atherosclerosis. Docosahexaenoic acid (DHA), a ω-3 polyunsaturated fatty acid (PUFA) has been shown to inhibit proliferation of several cell types implicating several different mechanisms. In the present study, we have investigated the effects of DHA on VSMC proliferation induced by stable and intermittent high glucose levels.

Confluent cultures of rat aortic VSMCs were treated with DHA for 24 hrs and then exposed to stable high glucose (25 mmol/L, SHG) or intermittent high glucose (5 mmol/L and 25 mmol/L alternating every 12 hrs, IHG) for 72 hrs. Cell proliferation was examined by the MTT viability assay, while apoptosis process was evaluated by the Hoechst staining, flow cytometry and caspase-3 activity assays.

Our data demonstrated that the hyper proliferation induced by stable and intermittent high glucose levels was significantly inhibited by the DHA pre-treatment. DHA significantly increased caspase-3 activity, resulting in enhanced DNA fragmentation and apoptosis.

Our results suggest that DHA reduced the high glucose-induced proliferation of VSMC and induced cell apoptosis.

• Apoptosis
• Diabetic atherosclerosis
• Docosahexaenoic acid
• Intermittent high glucose
• Proliferation
• Stable high glucose
• Vascular smooth muscle cells

• Angiogenesis Inhibitors/pharmacology
• Biological Transport
• Breast Neoplasms/genetics
• Breast Neoplasms/metabolism
• Cell Line, Tumor
• Docosahexaenoic Acids/pharmacology
• Endothelial Cells/drug effects
• Endothelial Cells/metabolism
• Exosomes/metabolism
• Female
• Gene Expression Regulation, Neoplastic/drug effects
• Gene Knockdown Techniques
• Humans
• MicroRNAs/genetics
• MicroRNAs/metabolism
• Neovascularization, Physiologic/drug effects
• Neovascularization, Physiologic/genetics
• Reproducibility of Results
• Signal Transduction/drug effects
• Vascular Endothelial Growth Factor A/metabolism
• rab GTP-Binding Proteins/genetics
• rab GTP-Binding Proteins/metabolism
• rab27 GTP-Binding Proteins

• P20 RR016478 NCRR NIH HHS
• U54 GM104938 NIGMS NIH HHS
• 3P20RR016478-09S2 NCRR NIH HHS

• Angiogenesis
• Angiopoietin 2
• Astrocyte
• Neuroprotection
• Omega-3 polyunsaturated fatty acids
• Stroke
• VEGF

Cancer associated fibroblasts (CAFs) are responsible for tumor growth, angiogenesis, invasion, and metastasis. Matrix metalloproteinase (MMP)-9 secreted from cancer stroma populated by CAFs is a prerequisite for cancer angiogenesis and metastasis. Omega-3 polyunsaturated fatty acids (omega-3 PUFA) have been reported to have anti-tumor effects on diverse types of malignancies. Fat-1 mice, which can convert omega-6 to omega-3 PUFA independent of diet, are useful to investigate the functions of endogenous omega-3 PUFA. To examine the effect of omega-3 PUFA on tumorigenesis, TC-1 cells, a murine epithelial cell line immortalized by human papillomavirus (HPV) oncogenes, were injected subcutaneously into fat-1 or wild type mice. Tumor growth and angiogenesis of the TC-1 tumor were significantly suppressed in fat-1 compared to wild type mice. cDNA microarray of the tumors derived from fat-1 and wild type mice revealed that MMP-9 is downregulated in fat-1 mice. Immunohistochemical study demonstrated immunoreactivity for MMP-9 in the tumor stromal fibroblasts was diffusely positive in wild type whereas focal in fat-1 mice. MMP-9 was expressed in primary cultured fibroblasts isolated from fat-1 and wild type mice but was not expressed in TC-1 cells. Co-culture of fibroblasts with TC-1 cells enhanced the expression and the proteinase activity of MMP-9, although the protease activity of MMP-9 in fat-1-derived fibroblasts was lower than that in wild type fibroblasts. Our data suggests that omega-3 PUFAs suppress MMP-9 induction and tumor angiogenesis. These findings may provide insight into mechanisms by which omega-3 PUFAs exert anti-tumor effects by modulating tumor microenvironment.

• Animals
• Cell Line, Transformed
• Cell Transformation, Neoplastic
• Coculture Techniques
• Disease Models, Animal
• Down-Regulation
• Fatty Acids, Omega-3/metabolism
• Fibroblasts/metabolism
• Gelatinases/metabolism
• Gene Expression Profiling
• Humans
• Matrix Metalloproteinase 9/genetics
• Matrix Metalloproteinase 9/metabolism
• Mice
• Neoplasms/genetics
• Neoplasms/metabolism
• Neoplasms/pathology
• Neovascularization, Pathologic
• Tumor Burden

Circulating levels of novel long-chain hydroxy fatty acids (called GTAs) were recently discovered in the serum of healthy subjects which were shown to be reduced in subjects with colorectal cancer (CRC), independent of tumor burden or disease stage. The levels of GTAs were subsequently observed to exhibit an inverse association with age in the general population. The current work investigates the biological activity of these fatty acids by evaluating the effects of enriched human serum extracts on cell growth and inflammation.

GTAs were extracted from commercially available bulk human serum and then chromatographically separated into enriched (GTA-positive) and depleted (GTA-negative) fractions. SW620, MCF7 and LPS stimulated RAW264.7 cells were treated with various concentrations of the GTA-positive and GTA-negative extracts, and the effects on cell growth and inflammation determined.

Enriched fractions resulted in poly-ADP ribose polymerase (PARP) cleavage, suppression of NFκB, induction of IκBα, and reduction in NOS2 mRNA transcript levels. In RAW264.7 mouse macrophage cells, incubation with enriched fractions prior to treatment with LPS blocked the induction of several pro-inflammatory markers including nitric oxide, TNFα, IL-1β, NOS2 and COX2.

Our results show that human serum extracts enriched with endogenous long-chain hydroxy fatty acids possess anti-inflammatory and anti-proliferative activity. These findings support a hypothesis that the reduction of these metabolites with age may result in a compromised ability to defend against uncontrolled cell growth and inflammation, and could therefore represent a significant risk for the development of CRC.

• Antineoplastic Agents/pharmacology
• Apoptosis/drug effects
• Breast Neoplasms/physiopathology
• Cell Line, Tumor
• Cell Proliferation/drug effects
• Curcumin/metabolism
• Curcumin/pharmacology
• Docosahexaenoic Acids/pharmacology
• Dose-Response Relationship, Drug
• Drug Synergism
• Female
• Gene Expression Profiling
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• PPAR gamma/metabolism
• Signal Transduction/drug effects
• Tumor Suppressor Protein p53/metabolism

• P30 CA023168 NCI NIH HHS
• R03CA137765-01A1 NCI NIH HHS

An organic extract from fresh shrimp (Litopenaeus vannamei) was studied for antimutagenic and antiproliferative properties using Salmonella typhimurium tester strains TA98 and TA100 with metabolic activation (S9) and a cancer cell line (B-cell lymphoma), respectively. Shrimp extract was sequentially fractionated by thin layer chromatography (TLC) and each fraction was tested for antimutagenic and antiproliferative activities. Crude organic extracts obtained from shrimp reduced the number of revertants caused by aflatoxina B₁, showing a dose-response type of relationship. Sequential TLC fractionation of the active extracts produced several antimutagenic and/or antiproliferative fractions. These results suggested that the lipid fraction of the tested species contained compounds with chemoprotective properties that reduce the mutagenicity of AFB₁ and proliferation of a cancer cell line.

• antimutagenicity
• antiproliferation
• cultured shrimp

The fish oil-derived ω-3 fatty acids, like docosahexanoic (DHA), claim a plethora of health benefits. We currently evaluated the antitumor effects of DHA, alone or in combination with cisplatin (CP) in the EAC solid tumor mice model, and monitored concomitant changes in serum levels of C-reactive protein (CRP), lipid peroxidation (measured as malondialdehyde; MDA) and leukocytic count (LC). Further, we verified the capacity of DHA to ameliorate the lethal, CP-induced nephrotoxicity in rats and the molecular mechanisms involved therein.

EAC-bearing mice exhibited markedly elevated LC (2-fold), CRP (11-fold) and MDA levels (2.7-fold). DHA (125, 250 mg/kg) elicited significant, dose-dependent reductions in tumor size (38%, 79%; respectively), as well as in LC, CRP and MDA levels. These effects for CP were appreciably lower than those of DHA (250 mg/kg). Interestingly, DHA (125 mg/kg) markedly enhanced the chemopreventive effects of CP and boosted its ability to reduce serum CRP and MDA levels. Correlation studies revealed a high degree of positive association between tumor growth and each of CRP (r = 0.85) and leukocytosis (r = 0.89), thus attesting to a diagnostic/prognostic role for CRP.

On the other hand, a single CP dose (10 mg/kg) induced nephrotoxicity in rats that was evidenced by proteinuria, deterioration of glomerular filtration rate (GFR, -4-fold), a rise in serum creatinine/urea levels (2–5-fold) after 4 days, and globally-induced animal fatalities after 7 days. Kidney-homogenates from CP-treated rats displayed significantly elevated MDA- and TNF-α-, but reduced GSH-, levels. Rats treated with DHA (250 mg/kg, but not 125 mg/kg) survived the lethal effects of CP, and showed a significant recovery of GFR; while their homogenates had markedly-reduced MDA- and TNF-α-, but -increased GSH-levels. Significant association was detected between creatinine level and those of MDA (r = 0.81), TNF-α ) r = 0.92) and GSH (r = -0.82); implying causal relationships.

DHA elicited prominent chemopreventive effects on its own, and appreciably augmented those of CP as well. The extent of tumor progression in various mouse groups was highly reflected by CRP levels (thus implying a diagnostic/prognostic role for CRP). Further, this study is the first to reveal that DHA can obliterate the lethal CP-induced nephrotoxicity and renal tissue injury. At the molecular level, DHA appears to act by reducing leukocytosis, systemic inflammation, and oxidative stress.

• Alkylating Agents/toxicity
• Animals
• Antibiotics, Antineoplastic/pharmacology
• Antioxidants/administration & dosage
• Dietary Supplements
• Docosahexaenoic Acids/administration & dosage
• Epirubicin/pharmacology
• Female
• Mammary Neoplasms, Experimental/blood supply
• Mammary Neoplasms, Experimental/drug therapy
• Mammary Neoplasms, Experimental/immunology
• Methylnitrosourea/toxicity
• Neovascularization, Pathologic/physiopathology
• Rats
• Rats, Sprague-Dawley
• Vascular Endothelial Growth Factor A/metabolism
• alpha-Tocopherol/administration & dosage

• Aldehydes/antagonists & inhibitors
• Aldehydes/toxicity
• Annexin A5/metabolism
• Apoptosis/drug effects
• Blotting, Western
• Cell Survival/drug effects
• Cells, Cultured
• Cytochromes c/metabolism
• Docosahexaenoic Acids/pharmacology
• Endothelial Cells/drug effects
• Flow Cytometry
• Glutathione/metabolism
• Humans
• Inhibitor of Apoptosis Proteins/antagonists & inhibitors
• Inhibitor of Apoptosis Proteins/biosynthesis
• Inhibitor of Apoptosis Proteins/genetics
• Oxidative Stress/drug effects
• RNA, Messenger/biosynthesis
• RNA, Small Interfering/pharmacology
• Reverse Transcriptase Polymerase Chain Reaction
• Stress, Physiological/pathology
• Superoxides/metabolism
• Transfection

Epidemiological evidence strongly links fish oil, which is rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), with low incidences of several types of cancer. The inhibitory effects of omega-3 polyunsaturated fatty acids on cancer development and progression are supported by studies with cultured cells and animal models. Propofol (2,6-diisopropylphenol) is the most extensively used general anesthetic–sedative agent employed today and is nontoxic to humans at high levels (50 μg/ml). Clinically relevant concentrations of propofol (3 to 8 μg/ml; 20 to 50 μM) have also been reported to have anticancer activities. The present study describes the synthesis, purification, characterization and evaluation of two novel anticancer conjugates, propofol-docosahexaenoate (propofol-DHA) and propofol-eicosapentaenoate (propofol-EPA).

The conjugates linking an omega-3 fatty acid, either DHA or EPA, with propofol were synthesized and tested for their effects on migration, adhesion and apoptosis on MDA-MB-231 breast cancer cells.

At low concentrations (25 μM), DHA, EPA or propofol alone or in combination had minimal effect on cell adhesion to vitronectin, cell migration against serum and the induction of apoptosis (only 5 to 15% of the cells became apoptotic). In contrast, the propofol-DHA or propofol-EPA conjugates significantly inhibited cell adhesion (15 to 30%) and migration (about 50%) and induced apoptosis (about 40%) in breast cancer cells.

These results suggest that the novel propofol-DHA and propofol-EPA conjugates reported here may be useful for the treatment of breast cancer.

• Antineoplastic Agents/toxicity
• Apoptosis/drug effects
• Breast Neoplasms/pathology
• Breast Neoplasms/physiopathology
• Caspases/metabolism
• Cell Adhesion/drug effects
• Cell Division/drug effects
• Cell Line, Tumor
• Cell Movement/drug effects
• Cell Survival/drug effects
• Docosahexaenoic Acids/toxicity
• Eicosapentaenoic Acid/toxicity
• Enzyme Activation/drug effects
• Female
• Free Radical Scavengers/toxicity
• HeLa Cells
• Humans
• Propofol/toxicity

• Animals
• Apoptosis/drug effects
• Colorectal Neoplasms/drug therapy
• Colorectal Neoplasms/metabolism
• Colorectal Neoplasms/pathology
• Cyclooxygenase 2
• Dinoprostone/metabolism
• Docosahexaenoic Acids/pharmacology
• Eicosapentaenoic Acid/pharmacology
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit
• Isoenzymes/metabolism
• Male
• Membrane Proteins
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Mitogen-Activated Protein Kinase 1/metabolism
• Mitogen-Activated Protein Kinase 3/metabolism
• Phosphorylation
• Prostaglandin-Endoperoxide Synthases/metabolism
• Signal Transduction
• Transcription Factors/metabolism
• Tumor Cells, Cultured/transplantation
• Vascular Endothelial Growth Factor A/metabolism

AIM: To study the effect of red oil A5 on pancreatic cancer cells and its possible mechanisms.

METHODS: Effect of different concentrations of red oil A5 on proliferation of three pancreatic cancer cell lines, AsPC-1, MiaPaCa-2 and S2013, was measured by ³H-methyl thymidine incorporation. Time-dependent effects of 1:32 000 red oil A5 on proliferation of three pancreatic cancer cell lines, were also measured by ³H-methyl thymidine incorporation, and Time-course effects of 1:32 000 red oil A5 on cell number. The cells were counted by Z1-Coulter Counter. Flow-cytometric analysis of cellular DNA content in the control and red oil A5 treated AsPC-1, MiaPaCa-2 and S2013 cells, were stained with propidium iodide. TUNEL assay of red oil A5-induced pancreatic cancer cell apoptosis was performed. Western blotting of the cytochrome c protein in AsPC-1, MiaPaCa-2 and S2013 cells treated 24 hours with 1:32 000 red oil A5 was performed. Proteins in cytosolic fraction and in mitochondria fraction were extracted. Proteins extracted from each sample were electrophoresed on SDS-PAGE gels and then were transferred to nitrocellulose membranes. Cytochrome c was identified using a monoclonal cytochrome c antibody. Western blotting of the caspase-3 protein in AsPC-1, MiaPaCa-2 and S2013 cells treated with 1:32 000 red oil A5 for 24 hours was carried out. Proteins in whole cellular lysates were electrophoresed on SDS-PAGE gels and then transferred to nitrocellulose membranes. Caspase-3 was identified using a specific antibody. Western blotting of poly-ADP ribose polymerase (PARP) protein in AsPC-1, MiaPaCa-2 and S2013 cells treated with 1:32 000 red oil A5 for 24 hours was performed. Proteins in whole cellular lysates were separated by electrophoresis on SDS-PAGE gels and then transferred to nitrocellulose membranes. PARP was identified by using a monoclonal antibody.

RESULTS: Red oil A5 caused dose- and time-dependent inhibition of pancreatic cancer cell proliferation. Propidium iodide DNA staining showed an increase of the sub-G0/G1 cell population. The DNA fragmentation induced by red oil A5 in these three cell lines was confirmed by the TUNEL assay. Furthermore, Western blotting analysis indicated that cytochrome c was released from mitochondria to cytosol during apoptosis, and caspase-3 was activated following red oil A5 treatment which was measured by procaspase-3 cleavage and PARP cleavage.

CONCLUSION: These findings show that red oil A5 has potent anti-proliferative effects on human pancreatic cancer cells with induction of apoptosis in vitro.

• Apoptosis/drug effects
• Caspase 3
• Caspases/metabolism
• Cell Count
• Cell Division/drug effects
• Cell Line, Tumor/cytology
• Cell Line, Tumor/drug effects
• Cytochromes c/metabolism
• Fish Oils/pharmacology
• Humans
• Pancreatic Neoplasms
• Poly(ADP-ribose) Polymerases/metabolism
• Thymidine/pharmacokinetics

• P50 CA072712 NCI NIH HHS
• CA72712 NCI NIH HHS

• Anticarcinogenic Agents/pharmacology
• Apoptosis/drug effects
• CCAAT-Enhancer-Binding Proteins/drug effects
• CCAAT-Enhancer-Binding Proteins/metabolism
• Caspases/drug effects
• Caspases/metabolism
• Cell Cycle/drug effects
• Cyclins/drug effects
• DNA Damage/drug effects
• DNA, Neoplasm/drug effects
• Flow Cytometry
• Humans
• Intracellular Signaling Peptides and Proteins
• Lipids/pharmacology
• Mitochondria/drug effects
• Mitochondria/pathology
• Mitochondria/physiology
• Pancreatic Neoplasms/genetics
• Pancreatic Neoplasms/pathology
• Proteins/drug effects
• Proteins/metabolism
• Transcription Factor CHOP
• Transcription Factors/drug effects
• Transcription Factors/metabolism
• Tumor Cells, Cultured
• GADD45 Proteins

• P50 CA072712 NCI NIH HHS
• CA72712 NCI NIH HHS