LncRNA ITGB2-AS1靶向miR-338-3p对肺癌细胞多西他赛耐药性的影响  

Effect of lncRNA ITGB2-AS1 on docetaxel resistance of lung cancer cells by targeting miR-338-3p

作　　者：杨士杰[1] 刘春盛[1] 高丽环 YANG Shi-jie;LIU Chun-sheng;GAO Li-huan(Department of Radiotherapy and Chemotherapy,Jizhong Energy Xingtai Mining Group General Hospital,Xingtai 054000,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：《实用药物与临床》2021年第1期17-22,共6页Practical Pharmacy and Clinical Remedies

基　　金：2011年邢台市科学技术研究与发展指导计划项目(2011ZC155)。

摘　　要：目的研究lncRNA ITGB2-AS1对肺癌细胞增殖、凋亡及多西他赛耐药性的影响及潜在的分子机制。方法用不同浓度(6.25μg/L、12.5μg/L、25μg/L、50μg/L、100μg/L、200μg/L)多西他赛(DTX)处理肺癌A549和多西他赛耐药细胞A549/DTX细胞,CCK8法测定DTX对A549细胞增殖抑制率和IC 50,qRT-PCR检测A549和A549/DTX细胞中lncRNA ITGB2-AS1和miR-338-3p的水平,Western blot检测细胞中CyclinD1、p21、Bax和Bcl-2表达水平,流式细胞术检测细胞凋亡率,双荧光素酶报告系统验证lncRNA ITGB2-AS1和miR-338-3p的靶向关系。结果多西他赛(DTX)(6.25μg/L、12.5μg/L、25μg/L、50μg/L、100μg/L、200μg/L)对A549/DTX细胞的抑制率显著低于A549细胞,呈浓度依赖性,A549/DTX对DTX的IC 50(256.80±10.22)μg/L显著高于A549细胞(23.57±2.11)μg/L;在A549/DTX中,lncRNA ITGB2-AS1含量显著高于A549细胞(P<0.05),miR-338-3p含量显著低于A549细胞(P<0.05);抑制lncRNA ITGB2-AS1联合12.5μg/L DTX处理可提高A549/DTX细胞凋亡率,增强DTX对A549/DTX细胞的增殖抑制率,上调p21和Bax水平,下调Cyclin D1和Bcl-2蛋白含量;lncRNA ITGB2-AS1靶向负调控miR-338-3p的表达;下调miR-338-3p可逆转抑制lncRNA ITGB2-AS1对A549/DTX细胞增殖、凋亡和多西他赛耐药性的作用。结论LncRNA ITGB2-AS1可靶向miR-338-3p调控A549/DTX细胞的增殖、凋亡及多西他赛耐药性。LncRNA ITGB2-AS1是肺癌潜在的分子靶点。Objective To investigate the effects of lncRNA ITGB2-AS1 on the proliferation,apoptosis and docetaxel resistance of lung cancer cells,as well as the potential molecular mechanism.Methods Lung cancer A549 cells and docetaxel resistant A549/DTX cells were treated with docetaxel(DTX)of different concentrations(6.25μg/L,12.5μg/L,25μg/L,50μg/L,100μg/L,200μg/L).CCK8 assay was used to determine the proliferation inhibition rate and IC 50 of DTX on A549 cells.qRT-PCR was applied to detect the levels of lncRNA ITGB2-AS1 and miR-338-3p in A549 and A549/DTX cells.Western blot was used to detect the expression levels of proteins CyclinD1,p21,Bax and Bcl-2.Flow cytometry was used to detect the cell apoptosis rate.Dual-luciferase reporter assay system was performed to examine the targeting relationship between ITGB2-AS1 and miR-338-3p.Results The inhibition rate of docetaxel(DTX)of 6.25μg/L,12.5μg/L,25μg/L,50μg/L,100μg/L,200μg/L on A549/DTX cells was significantly lower than that on A549 cells in a concentration-dependent manner.The IC 50(256.80±10.22)μg/L of A549/DTX on DTX was significantly higher than that on A549 cells(23.57±2.11)μg/L.In A549/DTX cells,the level of lncRNA ITGB2-AS1 was significantly higher than that in A549 cells(P<0.05),and the level of miR-338-3p was significantly lower than that in A549 cells(P<0.05).Inhibition of lncRNA ITGB2-AS1 combined with 12.5μg/L DTX treatment increased the apoptosis rate of A549/DTX cells,enhanced the proliferation inhibition rate of DTX on A549/DTX cells,up-regulated the level of p21 and Bax proteins,and down-regulated the level of CyclinD1 and Bcl-2.LncRNA ITGB2-AS1 targeted and negatively regulated the expression of miR-338-3p.Downregulation of miR-338-3p reversed the effects of lncRNA ITGB2-AS1 inhibition on proliferation,apoptosis and docetaxel resistance of A549/DTX cells.Conclusion LncRNA ITGB2-AS1 regulates the proliferation,apoptosis and docetaxel resistance of A549/DTX cells by targeting miR-338-3p.LncRNA ITGB2-AS1 is a potential molecular target for l

关 键 词：肺癌 LncRNA ITGB2-AS1 miR-338-3p 增殖 凋亡 多西他赛耐药性 

分 类 号：R734.2[医药卫生—肿瘤]