• biomarkers
• gastric carcinogenesis
• gastric carcinoma
• pathology
• pre-neoplastic lesions

• Humans
• Molecular Targeted Therapy
• Neoplasm Invasiveness
• Precancerous Conditions/pathology
• Risk Factors
• Stomach Neoplasms/classification
• Stomach Neoplasms/epidemiology
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology

• NET-2016-02363853 Italian Health Ministry and Veneto Region

Gastric carcinoma is the third leading cause of lethal cancer worldwide. Previous studies showed that Notch1 receptor intracellular domain (N1IC), the activated form of Notch1 receptor, promotes gastric cancer progression. It has been demonstrated that a significant cross-talk interplays between Notch pathways and microRNAs (miRNAs) in controlling tumorigenesis. This study identified an intronic microRNA-151 (miR-151), which consists of two mature miRNAs, miR-151-3p and miR-151-5p, as a Notch1 receptor-induced miRNA in gastric cancer cells. Activation of Notch1 pathway enhanced expressions of miR-151 and its host gene, focal adhesion kinase (FAK), in gastric cancer cells. The levels of miR-151 in gastric cancer samples were higher than those of adjacent non-tumor samples. Activated Notch1 pathway induced CBF1-dependent FAK promoter activity. The ectopic expression of miR-151 promoted growth and progression of SC-M1 gastric cancer cells including cell viability and colony formation, migration, and invasion abilities. Activated Notch1 pathway could augment progression of gastric cancer cells through miR-151-5p and FAK. The mRNA levels of pluripotency genes, Nanog and SOX-2, tumorsphere formation ability, tumor growth, and lung metastasis of SC-M1 cells were elevated by activated Notch1 pathway through miR-151-5p. Furthermore, miR-151-5p could target 3′-untranslated region (3′-UTR) of p53 mRNA and down-regulate p53 level in SC-M1 cells. Mechanistically, Notch1/miR-151-5p axis contributed to progression of SC-M1 cells through down-regulation of p53 which in turn repressed FAK promoter activity. Taken together, these results suggest that Notch1 pathway and miR-151-5p interplay with p53 in a reciprocal regulation loop in controlling gastric carcinogenesis.

• FAK
• Notch1 receptor
• gastric carcinogenesis
• miR-151
• p53

• Adenocarcinoma/genetics
• Adenocarcinoma/pathology
• Aged
• Biomarkers, Tumor/genetics
• Chromosomal Instability
• Chromosomes, Human, Pair 17/genetics
• DNA Copy Number Variations
• Esophageal Neoplasms/genetics
• Esophageal Neoplasms/pathology
• Female
• Humans
• Male
• Middle Aged
• Receptor, ErbB-2/genetics
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology

• AID
• DNA repair
• RNS
• ROS

• 8q24
• Gastric cancer
• MYC
• Survival
• TNFRSF11B

• Epigenetics
• Gastric cancer
• Genetics
• Helicobacter pylori
• Inflammation

Early detection and treatment of non-invasive neoplasms can effectively reduce the incidence of advanced gastric carcinoma (GC), but only when the lineage is continuous between non-invasive and advanced tumours. Although a fraction of non-invasive neoplasms progress to invasive GC, it is difficult to identify individual progression-prone non-invasive neoplasms. To classify non-invasive gland-forming gastric neoplasms into clusters of different levels of progression risk, we applied mucin phenotyping and genomic DNA microarray analyses to intramucosal gland-forming gastric neoplasms.

Formalin-fixed, paraffin-embedded tissues from 19 non-invasive and 24 invasive gland-forming neoplasms were obtained via endoscopic submucosal dissection or surgical excision. According to the Vienna classification, intramucosal neoplasms were classified as low-grade or high-grade non-invasive neoplasms (LGNs [category 3] and HGNs [category 4], respectively) or invasive carcinomas (intramucosal GCs and mucosal parts of submucosal or deeper GCs [category 5]). Neoplastic lesions were characterized by mucin phenotypes determined using monoclonal antibodies against MUC2, MUC5AC, MUC6, and CD10. Genomic DNA samples from mucosal neoplasms were subjected to array-based comparative genomic hybridization and subsequent unsupervised, hierarchical clustering with selected large-sized genes.

There was no significant difference in mucin phenotype between HGNs/LGNs and invasive carcinomas. The clustering classified samples into stable, unstable, and intermediate. The histological tumour grade or mucin phenotype of non-invasive neoplasms did not correlate with the clustering results. Each cluster may represent an independent lineage of different outcome because the size distribution of non-invasive tumours among the 3 clusters almost overlapped. In contrast, the unstable cluster alone included invasive carcinomas.

These findings suggest that the outcome of individual tumours is not stochastically determined but can be predicted from the genomic copy-number profile even at the non-invasive stage. Non-invasive neoplasms of the unstable clusters, which accounted for 21% of non-invasive neoplasms, may progress to invasive carcinomas, whereas those of stable cluster may not.

The online version of this article (doi:10.1186/s12920-015-0080-6) contains supplementary material, which is available to authorized users.

AIM: To investigate the differentiated whole genome expression profiling of gastric high- and low-grade intraepithelial neoplasia and early-stage adenocarcinoma.

METHODS: Gastric specimens from an upper magnifying chromoendoscopic targeted biopsy were collected from March 2010 to May 2013. Whole genome expression profiling was performed on 19 low-grade intraepithelial neoplasia (LGIN), 20 high-grade intraepithelial neoplasia (HGIN), 19 early-stage adenocarcinoma (EGC), and 19 chronic gastritis tissue samples using Agilent 4 × 44K Whole Human Genome microarrays. Differentially expressed genes between different types of lesions were identified using an unpaired t-test and corrected with the Benjamini and Hochberg false discovery rate algorithm. A gene ontology (GO) enrichment analysis was performed using the GeneSpring software GX 12.6. The differentially expressed gene was verified using a real-time TaqMan^® PCR assay with independent tissue samples, including 26 LGIN, 15 HGIN, 14 EGC, and 20 chronic gastritis. The expression of G₀S₂ were further validated by immunohistochemical staining (IHC) in 24 LGIN, 40 HGIN, 30 EGC and 61 chronic gastritis specimens.

RESULTS: The gene expression patterns of LGIN and HGIN tissues were distinct. There were 2521 significantly differentially expressed transcripts in HGIN, with 951 upregulated and 1570 downregulated. A GO enrichment analysis demonstrated that the most striking overexpressed transcripts in HGIN compared with LGIN were in the category of metabolism, defense response, and nuclear factor κB (NF-κB) cascade. While the vast majority of transcripts had barely altered expression in HGIN and EGC tissues, only 38 transcripts were upregulated in EGC. A GO enrichment analysis revealed that the alterations of the immune response were most prominent in the progression from HGIN to EGC. It is worth noting that, compared with LGIN, 289 transcripts were expressed at higher levels both in HGIN and EGC. A characteristic gene, G₀/G₁ switch 2 (G₀S₂) was one of the 289 transcripts and related to metabolism, the immune response, and the NF-κB cascade, and its expression was validated in independent samples through real-time TaqMan^® PCR and immunohistochemical staining. In real-time PCR analysis, the expression of G₀S₂ was elevated both in HGIN and EGC compared with that in LGIN (P < 0.01 and P < 0.001, respectively). In IHC analysis, G₀S₂ immunoreactivity was detected in the cytoplasmic of neoplastic cells, but was undetectable in chronic gastritis cells. The G₀S₂ expression in HGIN was higher than that of LGIN (P = 0.012, χ² = 6.28) and EGC (P = 0.008, χ² = 6.94).

CONCLUSION: A clear biological distinction between gastric high- and low-grade intraepithelial neoplasia was identified, and provides molecular evidence for clinical application.

• Gastric early-stage adenocarcinoma
• High-and low-grade intraepithelial neoplasia
• G₀/G₁ switch 2
• Whole genome expression microarray
• Quantitative real-time PCR
• Immunohistochemical staining

• Adenocarcinoma/chemistry
• Adenocarcinoma/metabolism
• Adenocarcinoma/pathology
• Adult
• Aged
• Biomarkers, Tumor/analysis
• Biomarkers, Tumor/genetics
• Biopsy
• Carcinoma in Situ/chemistry
• Carcinoma in Situ/genetics
• Carcinoma in Situ/pathology
• Cell Cycle Proteins/analysis
• Cell Cycle Proteins/genetics
• Computational Biology
• Databases, Genetic
• Diagnosis, Differential
• Female
• Gene Expression Profiling/methods
• Gene Expression Regulation, Neoplastic
• Genetic Predisposition to Disease
• Humans
• Immunohistochemistry
• Male
• Middle Aged
• Neoplasm Grading
• Neoplasm Staging
• Oligonucleotide Array Sequence Analysis
• Phenotype
• Predictive Value of Tests
• Real-Time Polymerase Chain Reaction
• Reproducibility of Results
• Reverse Transcriptase Polymerase Chain Reaction
• Stomach Neoplasms/chemistry
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology
• Transcription, Genetic

• Adenomatous Polyposis Coli Protein/genetics
• Chromosomes, Human, Pair 5/genetics
• DNA Copy Number Variations/genetics
• Genetic Association Studies
• Helicobacter pylori
• Humans
• Membrane Proteins
• Signal Recognition Particle
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology

Although mosaic autosomal chromosomal abnormalities are being increasingly detected as part of high-density genotyping studies, the clinical correlates are unclear. From an electronic medical record (EMR)–based genome-wide association study (GWAS) of peripheral arterial disease, log-R-ratio and B-allele-frequency data were used to identify mosaic autosomal chromosomal abnormalities including copy number variation and loss of heterozygosity. The EMRs of patients with chromosomal abnormalities and those without chromosomal abnormalities were reviewed to compare clinical characteristics. Among 3336 study participants, 0.75% (n = 25, mean age = 74.8 ± 10.7 years, 64% men) had abnormal intensity plots indicative of autosomal chromosomal abnormalities. A hematologic malignancy was present in 8 patients (32%), of whom 4 also had a solid organ malignancy while 2 patients had a solid organ malignancy only. In 50 age- and sex-matched participants without chromosomal abnormalities, there was a lower rate of hematologic malignancies (2% vs 32%, P < .001) but not solid organ malignancies (20% vs 24%, P = .69). We also report the clinical characteristics of each patient with the observed chromosomal abnormalities. Interestingly, among 5 patients with 20q deletions, 4 had a myeloproliferative disorder while all 3 men in this group had prostate cancer. In summary, in a GWAS of 3336 adults, 0.75% had autosomal chromosomal abnormalities and nearly a third of them had hematologic malignancies. A potential novel association between 20q deletions, myeloproliferative disorders, and prostate cancer was also noted.

• copy number variation
• genome-wide association studies
• loss of heterozygosity
• mosaic abnormalities
• mosaic deletion
• myeloproliferative disorders
• prostate cancer
• unipaternal disomy

Genistein has been shown to suppress the growth of several cancers through modulation of various pathways. However, the effects of genistein on the regulation of oncogenic microRNA-151 (miR-151) have not been reported. In this study, we investigated whether genistein could alter the expression of oncogenic miR-151 and its target genes that are involved in the progression and metastasis of prostate cancer (PCa). Real-time RT-PCR showed that the expression of miR-151 was higher in PC3 and DU145 cells compared with RWPE-1 cells. Treatment of PC3 and DU145 cells with 25 µM genistein down-regulated the expression of miR-151 compared with vehicle control. Inhibition of miR-151 in PCa cells by genistein significantly inhibited cell migration and invasion. In-silico analysis showed that several genes (CASZ1, IL1RAPL1, SOX17, N4BP1 and ARHGDIA) suggested to have tumor suppressive functions were target genes of miR-151. Luciferase reporter assays indicated that miR-151 directly binds to specific sites on the 3′UTR of target genes. Quantitative real-time PCR analysis showed that the mRNA expression levels of the five target genes in PC3 and DU145 were markedly changed with miR-151 mimics and inhibitor. Kaplan-Meier curves and log-rank tests revealed that high expression levels of miR-151 had an adverse effect on survival rate. This study suggests that genistein mediated suppression of oncogenic miRNAs can be an important dietary therapeutic strategy for the treatment of PCa.

• 3' Untranslated Regions/genetics
• Aged
• Antineoplastic Agents/pharmacology
• Base Sequence
• Cell Line, Tumor
• Cell Movement/drug effects
• Cell Movement/genetics
• Cell Proliferation/drug effects
• Computational Biology
• Disease Progression
• Gene Knockdown Techniques
• Genes, Reporter/genetics
• Genetic Vectors/genetics
• Genistein/pharmacology
• Humans
• Luciferases/genetics
• Male
• MicroRNAs/antagonists & inhibitors
• MicroRNAs/genetics
• Middle Aged
• Neoplasm Invasiveness
• Neoplasm Metastasis
• Oncogenes/genetics
• Prostatic Neoplasms/genetics
• Prostatic Neoplasms/pathology
• Up-Regulation/drug effects

• R01CA160079 NCI NIH HHS
• R01 CA160079 NCI NIH HHS
• R01CA138642 NCI NIH HHS
• T32 DK007790 NIDDK NIH HHS
• T32DK007790 NIDDK NIH HHS
• R01 CA138642 NCI NIH HHS

The relationship between benign uterine leiomyomas and their malignant counterparts, i.e. leiomyosarcomas and smooth muscle tumors of uncertain malignant potential (STUMP), is still poorly understood. The idea that a leiomyosarcoma could derive from a leiomyoma is still controversial. Recently MED12 mutations have been reported in uterine leiomyomas. In this study we asked whether such mutations could also be involved in leiomyosarcomas and STUMP oncogenesis. For this purpose we examined 33 uterine mesenchymal tumors by sequencing the hot-spot mutation region of MED12. We determined that MED12 is altered in 66.6% of typical leiomyomas as previously reported but also in 11% of STUMP and 20% of leiomyosarcomas. The mutated allele is predominantly expressed in leiomyomas and STUMP. Interestingly all classical leiomyomas exhibit MED12 protein expression while 40% of atypical leiomyomas, 50% of STUMP and 80% of leiomyosarcomas (among them the two mutated ones) do not express MED12. All these tumors without protein expression exhibit complex genomic profiles. No mutations and no expression loss were identified in an additional series of 38 non-uterine leiomyosarcomas. MED12 mutations are not exclusive to leiomyomas but seem to be specific to uterine malignancies. A previous study has suggested that MED12 mutations in leiomyomas could lead to Wnt/β-catenin pathway activation however our immunohistochemistry results show that there is no association between MED12 status and β-catenin nuclear/cytoplasmic localization. Collectively, our results show that subgroups of benign and malignant tumors share a common genetics. We propose here that MED12 alterations could be implicated in the development of smooth muscle tumor and that its expression could be inhibited in malignant tumors.

• Alleles
• Base Sequence
• Cell Transformation, Neoplastic/genetics
• Cell Transformation, Neoplastic/pathology
• DNA Mutational Analysis
• DNA, Complementary/genetics
• Female
• Gene Expression Regulation, Neoplastic
• Genome, Human/genetics
• Humans
• Leiomyoma/genetics
• Leiomyoma/pathology
• Mediator Complex/genetics
• Mediator Complex/metabolism
• Molecular Sequence Data
• Mutation/genetics
• Protein Transport
• Smooth Muscle Tumor/genetics
• Smooth Muscle Tumor/pathology
• Soft Tissue Neoplasms/genetics
• Soft Tissue Neoplasms/pathology
• Uterine Neoplasms/genetics
• Uterine Neoplasms/pathology
• Wnt Signaling Pathway/genetics
• beta Catenin/metabolism

• 3' Untranslated Regions
• Cell Line
• Cell Line, Tumor
• Cell Transformation, Neoplastic
• Cyclin-Dependent Kinase Inhibitor p21/antagonists & inhibitors
• Cyclin-Dependent Kinase Inhibitor p21/genetics
• Cyclin-Dependent Kinase Inhibitor p21/metabolism
• Down-Regulation
• Humans
• MicroRNAs/metabolism
• Neoplasms/genetics
• RNA, Messenger/metabolism
• Tumor Suppressor Protein p53/antagonists & inhibitors
• Up-Regulation

Genomic copy number aberrations (CNAs) in gastric cancer have already been extensively characterized by array comparative genomic hybridization (array CGH) analysis. However, involvement of genomic CNAs in the process of submucosal invasion and lymph node metastasis in early gastric cancer is still poorly understood. In this study, to address this issue, we collected a total of 59 tumor samples from 27 patients with submucosal-invasive gastric cancers (SMGC), analyzed their genomic profiles by array CGH, and compared them between paired samples of mucosal (MU) and submucosal (SM) invasion (23 pairs), and SM invasion and lymph node (LN) metastasis (9 pairs). Initially, we hypothesized that acquisition of specific CNA(s) is important for these processes. However, we observed no significant difference in the number of genomic CNAs between paired MU and SM, and between paired SM and LN. Furthermore, we were unable to find any CNAs specifically associated with SM invasion or LN metastasis. Among the 23 cases analyzed, 15 had some similar pattern of genomic profiling between SM and MU. Interestingly, 13 of the 15 cases also showed some differences in genomic profiles. These results suggest that the majority of SMGCs are composed of heterogeneous subpopulations derived from the same clonal origin. Comparison of genomic CNAs between SMGCs with and without LN metastasis revealed that gain of 11q13, 11q14, 11q22, 14q32 and amplification of 17q21 were more frequent in metastatic SMGCs, suggesting that these CNAs are related to LN metastasis of early gastric cancer. In conclusion, our data suggest that generation of genetically distinct subclones, rather than acquisition of specific CNA at MU, is integral to the process of submucosal invasion, and that subclones that acquire gain of 11q13, 11q14, 11q22, 14q32 or amplification of 17q21 are likely to become metastatic.

• Aged
• Aged, 80 and over
• Comparative Genomic Hybridization/methods
• Cortactin/metabolism
• DNA Copy Number Variations/genetics
• ErbB Receptors/metabolism
• Female
• Gastric Mucosa/metabolism
• Gastric Mucosa/pathology
• Genetic Heterogeneity
• Genome, Human/genetics
• Genomics
• Humans
• Immunohistochemistry
• Lymphatic Metastasis/pathology
• Male
• Middle Aged
• Models, Biological
• Neoplasm Invasiveness
• Receptor, ErbB-2/metabolism
• Sequence Deletion/genetics
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology