抑癌基因SPARCL1低表达对卵巢癌耐药和临床预后的影响  

Low expression of SPARCL1 contribute to cisplatin resistance and predict poor prognosis in ovarian cancer

作　　者：蔡香雪 李婷 韦露薇[2] 李力[2] 尹富强 Cai Xiangxue;Li Ting;Wei Luwei(Life Sciences Institute,Ministry of Education,Guangxi Medical University,Nanning530021;Key Laboratory of High-Incidence Tumor Prevention and Treatment,Ministry of Education,Guangxi Medical University,Nanning530021)

机构地区：[1]广西医科大学生命科学研究院,南宁530021 [2]广西医科大学区域性高发肿瘤早期防治研究教育部重点实验室,南宁530021

出　　处：《现代妇产科进展》2020年第4期256-262,共7页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金青年基金(No:81302283);广西自然科学基金回国基金(No:2014GXNSFCA118010);区域性高发肿瘤早期防治研究教育部重点实验室自主研究项目(GKE2015-ZZ18)资助。

摘　　要：目的:探讨SPARCL1表达对卵巢癌顺铂耐药的影响,阐明SPARCL1表达与耐药及预后的相关性。方法:RT-qPCR和Western blot法检测卵巢癌细胞中SPARCL1表达,免疫组化法(IHC)检测卵巢癌组织中SPARCL1表达,Kaplan-Meier生存曲线分析基因表达与无疾病生存期(DFS)和总生存期(OS)的关系。慢病毒转染过表达或干扰基因在顺铂耐药细胞SKOV3/DDP中的表达,CCK-8法检测细胞增殖并计算IC 50。转录组测序研究SPARCL1表达对卵巢癌细胞分子水平的影响。结果:与卵巢癌敏感组织相比,SPARCL1蛋白在耐药组织中显著低表达(P=0.001),且该蛋白低表达能预测卵巢癌不良DFS(P=0.001)和OS(P=0.021)。SPARCL1在顺铂耐药细胞SKOV3/DDP中显著下调表达,其过表达能减缓耐药细胞增殖速度并提高对顺铂的敏感性,而干扰其表达则能加快细胞增殖并降低细胞对顺铂的敏感性。转录组测序结果发现,SPARCL1过表达可能影响p53及细胞黏附分子等经典卵巢癌耐药调控通路。结论:SPARCL1低表达促进耐药且与不良OS和DFS显著相关,有望作为卵巢癌治疗靶标和潜在预后标记物应用于临床。Objective:To investigate the relationships of SPARCL1 with cisplatin resistance and prognosis in ovarian cancer.Methods:The expression of SPARCL1 was measured using RT-qPCR and Western blot for ovarian cancer cells,and using immunohistochemistry for tissues.The probability of survival and significance was calculated using the Kaplan-Meier analysis.The over-expression and interference expression of SPARCL1 in cisplatin-resistant SKOV3 cells(SKOV3/DDP)was performed using lentivirus-based over-expression vector and an RNA interference-based short hairpin RNA.Cell proliferation and IC 50 of ovarian cancer cells was determined by CCK-8 assay,and the effect of SPARCL1 on molecular changes of ovarian cancer cells was determined by transcriptome sequencing.Results:The protein expression of SPARCL1 was significantly lower in drug resistant ovarian cancer tissues than that in drug sensitive tissues(P=0.001).The low expression of SPARCL1 was significantly associated with poor disease-free survival(DFS)and overall survival(OS)(P=0.001 and 0.021,respectively).The mRNA expression of SPARCL1 was significantly decreased in SKOV3/DDP cells compared with SKOV3 cells.Using an over-expression vector and an RNA interference-based short hairpin RNA of SPARCL1,we found that up-regulation of SPARCL1 could significantly increase the sensitivity of the cells to cisplatin and inhibit the cell proliferation,while down-regulation of the gene indicated the opposite results.Transcriptome sequencing results indicated that the up-regulation of SPARCL1 was significantly correlated with p53 signaling pathway and cell adhesion molecules,which both were the typical pathways involved in the regulation of drug resistance in ovarian cancer.Conclusion:Down-regulation of SPARCL1 contributed to cisplatin resistance in ovarian cancer,and low expression of the gene predicted poor DFS and OS of ovarian cancer patients.SPARCL1 would be potentially used as a therapeutic target and a prognostic marker for clinical management of ovarian cancer.

关 键 词：SPARCL1 卵巢癌 顺铂耐药 预后 

分 类 号：R737.31[医药卫生—肿瘤]