内质网膜蛋白EMC1通过调控内质网应激诱导宫颈癌顺铂耐药的机制研究

顺铂耐药是宫颈癌治疗难题，但其具体机制不明。研究发现顺铂可诱导内质网应激，申请人前期通过宫颈癌顺铂耐药株和敏感株转录组测序，同时结合宫颈癌化疗数据库，发现高表达内质网膜蛋白EMC1与宫颈癌化疗耐药有关，且预示化疗患者不良预后；随后利用耐药株，通过体内外实验证实干扰EMC1可逆转宫颈癌顺铂耐药性；进一步通过蛋白聚集小体和免疫共沉淀等实验表明EMC1与分子伴侣GRP78相互作用，并激活适应性未折叠蛋白反应以缓解内质网应激。而其活化的适应性未折叠蛋白反应与肿瘤化疗抗性有关。因此，申请人推测EMC1可能通过招募分子伴侣GRP78促进错误折叠蛋白折叠以缓解内质网应激从而诱导宫颈癌顺铂耐药。本课题拟在前期基础上，1.阐明EMC1诱导宫颈癌耐药的具体分子机制；2.结合化疗样本分析EMC1的临床意义及作为宫颈癌化疗靶点的可能性，以期从内质网应激角度出发，为宫颈癌的化疗提供新的思路。

Cisplatin resistance is a disturbing problem for cervical cancer therapy, however its detailed mechanism is unclear. It’s known that cisplatin could induce endoplasmic reticulum stress (ERS), while its activated adaptive unfolded protein response (UPR) is supposed to be related with chemoresistance. Previously we conducted RNA sequencing between SiHa-parent (SiHa-Pa) and SiHa cisplatin resistance (SiHa-CR) cell lines, meanwhile, we combined with the chemotherapy database of cervical cancer (CC), finally we found out that endoplasmic reticulum protein 1(EMC1) was associated with chemoresistance and predicted poor survival in CC patients with chemotherapy; We then demonstrated that interfering EMC1 could reverse cisplatin resistance by SiHa-CR in vivo and in vitro. Mechanismly, we found that EMC1 interacted with molecular chaperone GRP78, and stimulated adaptive UPR to alleviate ERS by aggresome detection and co-immunoprecipitation assay. Therefore, we speculated that EMC1 may recruit molecular chaperone GRP78 to promote misfolded proteins fold and then alleviate ERS to induce cisplatin resistance in CC. Based on previous results, we aim to: 1) elucidate the molecular mechanism by which EMC1 promote CC chemoresistance, 2) analyze the clinical significance of EMC1 expression in chemoresistant tissues from CC patients, and determine the possibility of regarding EMC1 as a chemotherapeutic target. As a result, it will provide new ideas and solutions for improving the chemosensitivity in CC from the perspective of ERS.