MiR-495在GRP78介导的自噬参与胃癌多药耐药中的作用及机制的研究

项目名称： MiR-495在GRP78介导的自噬参与胃癌多药耐药中的作用及机制的研究

项目编号： No.81502402

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 赵国宏

作者单位： 中国人民解放军第四军医大学

项目金额： 17.5万元

中文摘要： 胃癌多药耐药是导致胃癌病人死亡的主要原因，其具体机制尚不清楚，miR-495已被发现参与调节多种肿瘤的发生发展，但其参与调节胃癌多药耐药的具体机制尚未阐明。前期我们发现miR-495在胃癌亲本细胞及耐药细胞中表达差异明显，并通过MTT，平板克隆、裸鼠成瘤实验、免疫化学染色等体内外实验验证其可能是通过调节自噬这条途径参与调节胃癌耐药细胞的生长及凋亡，为进一步研究其具体机制，我们通过生物信息学方法预测其下游靶基因并通过基因报告实验及免疫共沉淀对其进行验证，最后在胃癌临床标本中验证其表达及相关性，本研究想通过探讨miR-495-GRP78通过自噬参与调节胃癌多药耐药中的作用及机制，为临床胃癌多药耐药的发生发展提供一个新的治疗策略。

中文关键词： C07_胃肿瘤；miR-495；多药耐药；自噬；葡萄糖调节蛋白78

英文摘要： multi-drug resistance( MDR) is a major clinical obstacle in the treatment of gastric cancer (GC) and accounts for the majority of cancer-related mortality. MicroRNAs have been recently emerged as regulators of MDR through acting on multiple signaling pathways. We have identified miR-495 that diffential expressed between ADR GC cells using RT-PCR. And miR-495 showed the most strong effect on inhibiting GC cell autophagy and MDR. An MTT assay and xenograft model and Immunofluorescence staining demonstrated that the transfection of miR-495 induced autophagic activity and cell death in gastric cancer cells. We further investigated the underlying mechanism of mir-495 in GC autophagy and MDR. bioinformatics and proteomics will be used to predict and screen the potential target gene that MDR regulates. ChIP and Luciferase reporter assays will be used to identify the direct target gene. At last, expression of and mir-495 itself and target gene will be testified in human GC tissues. To reveal the mechanisms of miR-495-GRP78 contributes to autophagy-induced MDR in gastric cancer, and would be helpful in the therapeutic application to block GC MDR.

英文关键词： gastric cancer;miR-495;multi-drug resistance;autophagy;GRP78