CircZNF236/miR-214-3p/CHREBP通路在胃癌生长增殖中的作用及其机制

The abilities of growth and proliferation of gastric cancer play an irreplaceable role in the progression of gastric cancer. Our group found that the signal of CHREBP in gastric cancer was reduced when employing whole transcriptome resequencing. Additionally, in vitro experiments verified that CHREBP level was decreased in gastric cancer. Furthermore, reduced level of CHREBP might promote the growth and proliferation of gastric cancer via negatively regulating cyclin D1. Further analysis and utilization of whole transcriptome resequencing, bioinformatics and molecular biology indicated that circZNF236/miR-214-3p might regulate CHREBP. Thus，we put forward the assumption that high level of circZNF236 relieves the prohibited function of miR-214-3p on CHREBP level via sponging miR-214-3p, and then inhibits the growth and proliferation of gastric cancer. in vitro and in vivo experiments will be used to explore the role of CHREBP in growth and proliferation of gastric cancer, RIP, CHIP and others assays will be utilized to discover the upstream and downstream regulation of CHREBP. The aim of this.study is to explore the function, mechanism and clinical significance of circZNF236/miR-214-3p/CHREBP axis in the growth and proliferation of gastric cancer and to provide novel biomarkers for diagnosis and prognosis of gastric cancer.

胃癌细胞生长增殖能力影响胃癌的发生发展。课题组通过全转录组测序发现CHREBP在胃癌中低表达，体外实验进一步证实CHREBP在胃癌组织中低表达；低表达的CHREBP可能通过负向调控cyclin D1，促进胃癌生长增殖。进一步分析并利用全转录组测序、生物信息学、分子生物学，我们发现circZNF236/miR-214-3p可能调控CHREBP。因此推测circZNF236通过海绵样作用吸附miR-214-3p，影响CHREBP的表达，从而抑制胃癌生长增殖。本项目拟采用体内外实验研究CHREBP在胃癌生长增殖中的作用，RIP、CHIP等实验研究CHREBP的上、下游调控机制。本研究旨在阐明circZNF236/miR-214-3p/CHREBP通路在胃癌生长增殖中的作用、机制及其临床意义，期望为胃癌诊治和预后判断提供新的分子靶标。

胃癌是人类最常见的恶性肿瘤之一，在全球范围内具有较高的发病率和死亡率。在我国，胃癌的发病率和死亡率均位居第2位，每年新发病例约68万人，死亡病例约50万人，严重威胁大众身体健康。课题组前期发现CHREBP在胃癌中低表达，与胃癌的生长增殖密切相关，且低表达的CHREBP可能通过负向调控cyclinD1，促进胃癌生长增殖。因此，在本课题研究中，首先利用体内外实验明确CHREBP表达改变对胃癌细胞生长、增殖及细胞周期的影响，利用WB等分子生物学技术探讨CHREBP调控cyclin D1影响胃癌的生长增殖的机制研究。研究结果显示：1.公共数据库Oncomine提示CHREBP在胃癌中低表达，免疫组化和Qrt-pcr结果亦提示CHREBP在胃癌组织中低表达，且与T分期，神经侵犯，TNM分期负相关。Kaplan-Meier和Cox回归分析结果提示CHREBP表达与预后密切相关，可能作为胃癌预后独立的影响因子。2.CCK8、Edu、平板克隆、细胞周期、细胞凋亡等体外细胞实验发现下调CHREBP表达后，可以促进胃癌细胞的生长增殖，抑制其凋亡；上调CHREBP表达后，可以抑制胃癌细胞的生长增殖，促进其凋亡。体内动物实验也发现，上调CHREBP表达后，可抑制肿瘤的生长增殖。3.免疫荧光、双荧光素酶报告基因实验、WB等分子生物学实验提示CHREBP可以靶向调控CyclinD1的表达，影响下游Rb-E2F1分子通路，进而影响胃癌细胞的生物学功能。该研究明确CHREBP在胃癌生长增殖中的作用；阐明CHREBP对cyclinD1的负向调控机制；揭示了CHREBP/cyclinD1/Rb/E2F1通路在胃癌生长增殖的具体分子机制，为揭示胃癌的发生发展提供新的研究思路。