miR-424-5p靶向ARK5逆转胃癌耐药细胞对顺铂的耐药性的影响及机制研究  

Study on influence and mechanism of miR-424-5p targeting ARK5 reversing cisplatin resistance in drug-resistant gastric cancer cells

作　　者：杨光[1] 武鸣霞 YANG Guang;WU Mingxia(Tieling Central Hospital,Tieling 112000,Liaoning Province,China)

机构地区：[1]铁岭市中心医院,辽宁铁岭112000

出　　处：《世界临床药物》2021年第12期1073-1080,共8页World Clinical Drug

基　　金：辽宁省科学技术厅重点研发计划(2017JM126)。

摘　　要：目的探讨miR-424-5p对胃癌细胞顺铂(cisplatin,DDP)耐药的影响及其作用机制。方法利用RT-qPCR检测胃癌DDP耐药细胞(SGC7901/DDP)及其亲代细胞(SGC7901)中miR-424-5p及ARK5的表达水平;利用Lipofectamine 2000将miR-424-5p mimic、siR-ARK5、miR-424-5p mimic+pcDNA-ARK5转染至SGC7901/DDP中,通过MTT法测定各转染组SGC7901/DDP细胞对DDP耐药性的影响;以Target Scan预测miR-424-5p的潜在靶基因,采用双荧光素酶试验进行验证;利用免疫印迹(western blot,WB)试验测定转染后ARK5蛋白的表达水平。结果胃癌DDP耐药细胞中miR-424-5p表达显著下调,ARK5表达显著上调;MTT试验显示,过表达miR-424-5p及干扰ARK5表达降低了DDP耐药细胞的耐药性,共同转染miR-424-5p mimic和pcDNA-ARK5逆转了miR-424-5p mimic单独转染对胃癌细胞耐药性的影响;Target Scan预测发现,ARK5是miR-424-5p的潜在靶基因,双荧光素酶试验证实ARK5是miR-424-5p的靶基因;Western-blot试验结果显示miR424-5p负向调控ARK5蛋白的表达。结论胃癌DDP耐药细胞中miR-424-5p表达下调,ARK5表达上调;miR-424-5p通过靶向抑制ARK5表达有效逆转胃癌DDP耐药细胞的耐药性。Objective To investigate the effects of miR-424-5p on cisplatin(DDP)resistance of gastric cancer cells and its mechanism.Methods RT-qPCR was used to determine the expression of miR-424-5p and ARK5 in DDP-resistant gastric cancer cells(SGC7901/DDP)and its parent cells(SGC7901).Lipofectamine 2000 was used to transfect miR-424-5p mimic,siR-ARK5,miR-424-5p mimic+PCDNA-ARK5 into SGC7901/DDP.MTT assay was used to determine the effects of SGC7901/DDP cells on DDP resistance in each transfection group.The potential target genes of miR-424-5p were predicted by Target Scan and verified by dual luciferase assay.Western blot was used to determine the expression level of ARK5 protein after transfection.Results The miR-424-5p expression was significantly down-regulated and ARK5 expression was significantly up-regulated in DDP-resistant gastric cancer cells.MTT assay showed that overexpression of miR-424-5p and interference of ARK5 expression reduced the drug resistance of DDP-resistant cells.Co-transfection of miR-424-5p mimic and PCDNA-ARK5 reversed the effects of single transfection of miR-424-5p mimic on drug resistance of gastric cancer cells.Target Scan predicted that ARK5 was the potential target gene of miR-424-5p,and dual luciferase assay confirmed that ARK5 was the target gene of miR-424-5p.Western blot showed that miR-424-5p negatively regulated the expression of ARK5 protein.Conclusion MiR-424-5p expression was down-regulated and ARK5 expression was up-regulated in DDPresistant gastric cancer cells.MiR-424-5p effectively reversed drug resistance of DDP-resistant gastric cancer cells by targeting inhibition of ARK5 expression.

关 键 词：胃癌 miR-424-5p ARK5 顺铂 耐药性 

分 类 号：R-3[医药卫生] R979.11