miR-129-5p靶向LARP1基因调控肺癌细胞顺铂耐药  

miR-129-5p regulates cisplatin resistance of lung cancer cells by targeting LARP1

作　　者：杨士杰[1] 刘春盛[1] 高丽环 赵庆书 刘孟奇[1] YANG Shijie;LIU Chunsheng;GAO Lihuan;ZHAO Qingshu;LIU Mengqi(Department of Radiotherapy and Chemotherapy,Jizhong Energy Xingtai Mining Group General Hospital,Xingtai 054000,Hebei,China)

机构地区：[1]冀中能源邢台矿业集团总医院放化疗科,河北邢台054000

出　　处：《西部医学》2020年第9期1298-1303,共6页Medical Journal of West China

基　　金：邢台市科学技术研究与发展指导计划项目(2011ZC155)。

摘　　要：目的探讨miR-129-5p能否通过调控La相关蛋白1(LARP1)表达,从而调节肺癌细胞对顺铂(DDP)的敏感性。方法通过实时荧光定量PCR(qRT-PCR)和蛋白质印记(Western blot)检测肺癌细胞A549和肺癌顺铂耐药细胞A549/DDP中miR-129-5p、LARP1以及多药耐药相关蛋白1(MRP1)的表达水平。将A549/DDP分为对照组(NC)、miR-con组、miR-129-5p组、si-con组和si-LARP1组、miR-129-5p+pcDNA-con组、miR-129-5p+pcDNA-LARP1组。细胞计数试剂盒(CCK-8)检测细胞存活率和对顺铂的半数抑制浓度(IC50)。Western blot检测细胞周期蛋白D1(CyclinD1)和MRP1蛋白表达。双荧光素酶报告基因实验和Western blot检测验证miR-129-5p和LARP1的靶向调控关系。结果与A549细胞相比,A549/DDP细胞中miR-129-5p的表达水平降低,LARP1和MRP1的表达水平升高,差异均有统计学意义(P<0.05)。与miR-con组比较,miR-129-5p组A549/DDP细胞存活率、IC50、CyclinD1和MRP1蛋白表达均降低,差异均有统计学意义(P<0.05)。与si-con组比较,si-LARP1组A549/DDP细胞存活率、IC50降低,CyclinD1和MRP1蛋白表达降低,差异均有统计学意义(P<0.05)。与miR-129-5p+pcDNA-con组比较,miR-129-5p+pcDNA-LARP1组A549/DDP细胞存活率、IC50升高,CyclinD1和MRP1蛋白表达升高,差异均有统计学意义(P<0.05)。LARP1是miR-129-5p的靶基因。过表达miR-129-5p后LARP1蛋白表达降低,沉默miR-129-5p后LARP1蛋白表达升高,差异均有统计学意义(P<0.05)。结论miR-129-5p通过靶向下调LARP1表达抑制肺癌细胞增殖,提高肺癌细胞的顺铂敏感性。Objective To investigate whether miR-129-5p could regulate the sensitivity of lung cancer cells to cisplatin(DDP)by regulating the expression of La-related protein 1(LARP1).Methods The expression levels of miR-129-5p,LARP1 and multidrug resistance-associated protein(MRP1)in lung cancer cells A549 and lung cancer cisplatin-resistant cells A549/DDP were detected by real-time quantitative PCR and Western blot.A549/DDP cells were divided into control(NC),miR-con,miR-129-5P,si-con,si-LARP1,miR-129-5p+pcDNA-con and miR-129-5p+pcDNA-LARP1 groups.The cell counting kit(CCK-8)was used to detect cell viability and the half-inhibitory concentration(IC50)against cisplatin.Western blot detected the expression levels of CyclinD1 and MRP1 proteins.The dual luciferase reporter gene assay and Western blot assay were used to verify the targeted and regulatory relationship between miR-129-5p and LARP1.Results Compared with A549 cells,the expression level of miR-129-5p was decreased in A549/DDP cells,while the expression levels of LARP1 and MRP1 were increased,the difference was statistically significant(P<0.05).Compared with miR-con group,A549/DDP the cell survival rate and IC50 of A549/DDP cells in miR-129-5p group were decreased,and expression of CyclinD1 and MRP1 proteins were decreased,the difference was statistically significant(P<0.05).Compared with the si-con group,the survival rate and IC50 of A549/DDP cells in the si-LARP1 group were decreased,and the expression of CyclinD1 and MRP1 proteins were decreased,the difference was statistically significant(P<0.05).Compared with miR-129-5p+pcDNA-con group,the survival rate and IC50 of A549/DDP cells in the miR-129-5p+pcDNA-LARP1 group were increased,and the expression of CyclinD1 and MRP1 proteins were increased,the difference was statistically significant(P<0.05).LARP1 was the target gene of miR-129-5p.LARP1 protein expression was decreased after over-expressing miR-129-5p,whereas LARP1 protein expression was increased after silencing miR-129-5p,the difference was statistically s

关 键 词：肺癌细胞 miR-129-5p LARP1 顺铂耐药 敏感性 

分 类 号：R734.2[医药卫生—肿瘤]