Magic™ Anti-c-Myc polyclonal antibody

Simple Summary Esophageal adenocarcinoma has become a major clinical challenge in the western world due to its rapid increasing incidence and poor overall prognosis. Understanding the molecular events of its tumorigenesis is the key to better diagnosis and development of better therapeutic strategies. In the current study we aimed to identify epigenetic alteration targets in esophageal adenocarcinoma. We focused on a candidate gene,NDRG4(N-myc downregulated gene 4). We found thatNDRG4was frequent downregulated in esophageal adenocarcinoma through DNA hypermethylation of its promoter region. Re-expression of NRDG4 in cancer cells significantly suppressed tumor growth via inhibition of cell proliferation. These results will improve our understanding on how dysfunction ofNDRG4contributes to esophageal adenocarcinoma. DNA hypermethylation ofNDRG4may be a useful biomarker in clinical monitoring of esophageal adenocarcinoma patients. The incidence of esophageal adenocarcinoma (EAC) has been rising dramatically in the past few decades in the United States and Western world. The N-myc downregulated gene 4 (NDRG4) belongs to the human NDRG family. In this study, we aimed to identify the expression levels, regulation, and functions ofNDRG4in EAC. Using an integrative epigenetic approach, we identified genes showing significant downregulation in EAC and displaying upregulation after 5-Aza-deoxycitidine. Among these genes, likely to be regulated by DNA methylation,NDRG4was among the top 10 candidate genes. Analyses of TCGA (The Cancer Genome Atlas) and GEO (Gene Expression Omnibus) data sets and EAC tissue samples demonstrated thatNDRG4was significantly downregulated in EAC (p< 0.05). Using Pyrosequencing technology for quantification of DNA methylation, we detected thatNDRG4promoter methylation level was significantly higher in EAC tissue samples, as compared to normal esophagus samples (p< 0.01). A strong inverse correlation betweenNDRG4methylation and its gene expression levels (r= -0.4,p< 0.01) was observed. Treatment with 5-Aza restored theNDRG4expression, confirming that hypermethylation is a driving force forNDRG4silencing in EAC. Pathway and gene set enrichment analyses of TCGA data suggested thatNDRG4is strongly associated with genes related to cell cycle regulation. Western blotting analysis showed significant downregulation of Cyclin D1, CDK4 and CDK6 in EAC cells after overexpression of NDRG4. Functionally, we found that the reconstitution of NDRG4 resulted in a significant reduction in tumor cell growth in two-dimensional (2D) and three-dimensional (3D) organotypic culture models and inhibited tumor cell proliferation as indicated by the EdU (5-ethynyl-2 '-deoxyuridine) proliferation assay.

Somatic copy number alterations (CNA) are common in endometrial serous carcinoma (ESC). We used the Tumor Cancer Genome Atlas Pan Cancer dataset (TCGA Pan Can) to explore the impact of somatic CNA and gene expression levels (mRNA) of cancer-related genes in ESC. Results were correlated with clinico-pathologic parameters such as age of onset, disease stage, progression-free survival (PFS) and overall survival (OS) (n = 108). 1,449 genes with recurrent somatic CNA were identified, observed in 10% or more tumor samples. Somatic CNA and mRNA expression levels were highly correlated (r> = 0.6) for 383 genes. Among these, 45 genes were classified in the Tier 1 category of Cancer Genome Census-Catalogue of Somatic Mutations in Cancer. Eighteen of 45 Tier 1 genes had highly correlated somatic CNA and mRNA expression levels includingARNT,PIK3CA,TBLXR1,ASXL1,EIF4A2,HOOK3,IKBKB,KAT6A,TCEA1,KAT6B,ERBB2,BRD4,KEAP1,PRKACA,DNM2,SMARCA4,AKT2,SS18L1. Our results are in agreement with previously reported somatic CNA forERBB2,BRD4andPIK3Cin ESC. In addition,AKT2(p = 0.002) andKAT6A (p = 0.015) amplifications were more frequent in tumor samples from younger patients (<60), andCEBPA(p = 0.028) andMYC(p = 0.023) amplifications were more common with advanced (stage III and IV) disease stage. Patients with tumors carryingKAT6AandMYCamplifications had shorter PFS and OS. The hazard ratio (HR) ofKAT6Awas 2.82 [95 CI 1.12-7.07] for PFS and 3.87 [95 CI 1.28-11.68] for OS. The HR ofMYCwas 2.25 [95 CI 1.05-4.81] and 2.62[95 CI 1.07-6.41] for PFS and OS, respectively.