Screening and validation of markers related to diagnosis and prognosis assessment of gastric cancer based on miRNA-mRNA network and their potential molecular mechanisms / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：通过生物信息学方法探索并实验验证胃癌相关标志物miR-1-3p对胃癌细胞增殖的作用及其分子机制。方法：收集TCGA数据库中胃癌（n=375）及癌旁组织（n=45）的转录组数据，构建胃癌特异性mRNA-miRNA网络，筛选潜在的miRNA类标志物，利用TargetScan预测标志物的下游靶基因且分析它们的功能。选取人正常胃上皮细胞GES-1及胃癌细胞AGS、MKN45、NCI-N87，用qPCR法检测细胞中miR-1-3p和心肌蛋白（MYOCD）的表达，用lipofectamine 2000将miR-1-3p模拟物转染至胃癌细胞中，CCK-8法测定轨染后细胞的增殖能力，WB法测定MYOCD的表达量，双荧光素酶报告基因实验验证miR-1-3p与MYOCD之间的靶向结合关系。结果：通过数据库数据分析得到差异表达的259个miRNA和7 545个mRNA，构建胃癌特异性mRNA-miRNA调节网络，分析网络中脆弱结构后确定miR-1-3p为潜在的胃癌标志物，ROC曲线和Kaplan-Meier分析显示其对胃癌的诊断和预后评估有重要意义。细胞实验显示miR-1-3p在胃癌细胞中呈低表达（P<0.05），过表达miR-1-3p可抑制胃癌细胞AGS和MKN-45的增殖能力（P<0.05或P<0.01），且可抑制MYOCD的表达（P<0.01）。TargetScan数据库预测到MYOCD的3'UTR区域中有两个与miR-1-3p结合的位点，双荧光素酶报告基因实验证实miR-1-3p与MYOCD靶向结合且负调控MYOCD的表达（P<0.01）。结论: miR-1-3p可能是胃癌诊断和预后相关潜在的标志物，且miR-1-3p可能是通过靶向MYOCD来影响胃癌细胞的增殖。

Identification and functional analysis of pathogenic genes and key transcription factors in prostate adenocarcinoma / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the pathogenesis of prostate cancer by analyzing the associated hub gene modules of prostate cancer and identifying key transcription factors and genes that affect these modules. Methods: WGCNA (weighted gene co-expressed network analysis) was used to identify hub gene modules associated with important clinicopathological features of prostate cancer, such as pathological staging, Gleason grading etc. The OPOSSUM online tool was used to analyze the transcription factors enriching and regulating those genes. Pathway enrichment analysis and protein-protein interaction network analysis were used to identify key genes in prostate cancer. Finally, the effects of these genes on clinical features and disease-free survival (DFS) of prostate cancer patients were analyzed. Results: Three hub modules were identified, and they were highly associated with pathologic T stage, pathologic N stage and Gleason grading of prostate cancer, respectively. Further screening revealed 13 key dysregulated transcription factors that participated in the regulation of these three hub modules. The differentially expressed genes regulated by the 13 key transcription factors were significantly enriched in Calcium signaling pathway, cGMP-PKG signaling pathway and cAMP signaling pathway. 14 key genes (PRKG1, PRKG2, CYSLTR2, GRPR, CHRM3, ADCY5, ADRA1D, EDNRA, EDNRB, CYSLTR2, AGTR1, GRPR, GRIA1 and OXT) were at important nodes in the gene network. Among them, the high expression of ADRA1A, PRKG2, CHRM3, ADRA1D and EDN3 significantly extended the DFS of patients with prostate cancer (all P<0.01). Conclusion: ADRA1A, PRKG2, CHRM3, ADRA1D and EDN3 are regulated by key dysregulated transcription factors and highly associated with clinical features of prostate cancer. Their high expressions will significantly prolong the DFS of prostate cancer patients, which may shed light to the discovery of mechanism in prostate adenocarcinoma.