微小RNA-199a-5p对非小细胞肺癌紫杉醇耐药性的影响及其机制  

Effect of microRNA-199a-5p on Paclitaxel Resistance in Non-small Cell Lung Cancer and Its Mechanism

作　　者：蒙冲[1] 刘礼荣[1] 刘凯[1] MENG Chong;LIU Lirong;LIU Kai(Department of Respiratory and Critical Care Medicine,Hainan Provincial People's Hospital,Haikou 570311,China)

机构地区：[1]海南省人民医院呼吸与危重症医学科,海口570311

出　　处：《医药导报》2023年第5期632-638,共7页Herald of Medicine

基　　金：海南省卫生健康行业科研项目(20A200127)。

摘　　要：目的 探讨微小RNA-199a-5p(miR-199a-5p)对非小细胞肺癌(NSCLC)细胞紫杉醇(PTX)耐药性的影响及其机制。方法 采用实时荧光定量聚合酶链反应(qRT-PCR)分析人NSCLC细胞株A549和耐药NSCLC细胞株A549/PTX中miR-199a-5p相对表达水平;采用细胞增殖试剂盒(CCK-8)分析A549和A549/PTX细胞对PTX的耐药性;双荧光素酶实验验证miR-199a-5p与热休克因子-1(HSF-1)的靶向关系。采用CCK-8和TUNEL分别检测细胞增殖、凋亡情况;采用Western blotting检测细胞中HSF-1、热休克蛋白(HSP)90、HSP60、P-糖蛋白(P-gp)蛋白表达水平。结果 A549/PTX细胞对PTX的耐药性显著高于A549细胞(P<0.05),且A549/PTX细胞对PTX的半数抑制浓度(IC_(50))高于A549细胞。耐药细胞株A549/PTX中miR-199a-5p表达显著下调,HSF-1 mRNA显著上调(P<0.05)。在耐药细胞株A549/PTX中miR-199a-5p负靶向调控HSF-1表达(P<0.05)。在A549/PTX细胞中过表达miR-199a-5p或抑制HSF-1可降低PTX的IC_(50)值,促进细胞凋亡,下调HSF-1、HSP90、HSP60和P-gp蛋白水平(P<0.05);而过表达HSF-1则可在一定程度上逆转上调miR-199a-5p对A549/PTX细胞的影响(P<0.05)。结论 miR-199a-5p通过抑制HSF-1减弱NSCLC细胞PTX耐药性。Objective To investigate the effect and its mechanism of microRNA-199a-5p(miR-199a-5p)on the resistance of non-small cell lung cancer(NSCLC)cells to paclitaxel(PTX).Methods The relative expression levels of miR-199a-5p in human NSCLC cell line A549 and drug-resistant NSCLC cell line A549/PTX were detected by qRT-PCR;the resistance of A549 and A549/PTX cells to PTX were detected by CCK-8.The targeting relationship between miR-199a-5p and heat shock factor-1(HSF-1)was verified by a dual luciferase experiment.Cell proliferation and apoptosis were detected by CCK-8 and TUNEL,respectively;the expression levels of HSF-1,heat shock protein(HSP)90,HSP60,and P-glycoprotein(P-gp)proteins of cells were detected by Western blotting.Results The resistance of A549/PTX cells to PTX was significantly higher than that of A549 cells(P<0.05),and the IC 50 of A549/PTX cells to PTX was higher than that of A549 cells.The expression of miR-199a-5p in the drug-resistant cell line A549/PTX was significantly down-regulated,and HSF-1 mRNA was significantly up-regulated(P<0.05).In the drug-resistant cell line A549/PTX,miR-199a-5p negatively targeted the expression of HSF-1(P<0.05).Overexpression of miR-199a-5p or inhibition of HSF-1 in A549/PTX cells could reduce PTX IC 50 values,promote cell apoptosis,down-regulate HSF-1,HSP90,HSP60,and P-gp protein levels(P<0.05);overexpression of HSF-1 could reverse the effect of up-regulation of miR-199a-5p on A549/PTX cells to a certain extent(P<0.05).Conclusion MiR-199a-5p can attenuate the PTX resistance of NSCLC cells by inhibiting HSF-1.

关 键 词：紫杉醇 微小RNA-199a-5p 非小细胞肺癌 耐药性 热休克因子-1 

分 类 号：R979.1[医药卫生—药品] R734.2[医药卫生—药学]