miR-329靶向调控细胞周期相关转录因子基因逆转非小细胞肺癌A549细胞顺铂耐药的机制研究  

Mechanisms of miR-329 targeting the cell cycle related transcription factor gene to reverse cisplatin resistance in non-small cell lung cancer A549 cells

作　　者：刘鑫[1] 李汉杰[3] 陈子溦 LIU Xin;LI Han-jie;CHEN Zi-wei(School of Nursing and Rehabilitation,Xi’an Medical College,Xi’an 710021,Shaanxi Province,China;Clinical Medical College,Xi’an Medical College,Xi’an 710021,Shaanxi Province,China;Department of Thoracic Surgery,The Second Affiliated Hospital of Xi’an Medical College,Xi’an 710038,Shaanxi Province,China)

机构地区：[1]西安医学院护理与康复学院,陕西西安710021 [2]西安医学院临床医学院,陕西西安710021 [3]西安医学院第二附属医院胸外科,陕西西安710038

出　　处：《中国临床药理学杂志》2022年第22期2688-2692,共5页The Chinese Journal of Clinical Pharmacology

基　　金：陕西省科技厅自然科学基础研究计划基金资助项目(2019JQ-521)。

摘　　要：目的探究miR-329靶向调控细胞周期相关转录因子(E2F1)基因对非小细胞肺癌A549细胞顺铂(DDP)耐药的影响。方法将DDP耐药细胞株A549/DDP细胞分为空白组(细胞未经任何处理)、对照组(转染miR-329-mimics-NC)和实验组(转染miR-329-mimics)。用实时荧光定量聚合酶链反应法测定miR-329和E2F1 mRNA的表达水平,用细胞计数试剂盒-8法检测细胞增殖情况,用蛋白质印迹法检测E2F1蛋白、重组人叉头框蛋白L2(FOXL2)和整合素α2(ITGA2)蛋白的表达情况。结果A549细胞和A549/DDP细胞的miR-329相对表达水平分别为1.00±0.16和0.53±0.08,差异有统计学意义(P<0.05)。实验组、对照组和空白组的miR-329相对表达水平分别为1.68±0.26,1.02±0.16和1.00±0.15,E2F1 mRNA相对表达水平分别为0.33±0.05,1.01±0.15和1.00±0.16,细胞增殖抑制率分别为(28.95±4.33)%,(14.03±2.10)%和(13.26±1.98)%,半抑制浓度分别为(28.40±1.72),(47.85±5.71)和(45.63±3.18)μmol·L^(-1),E2F1蛋白相对表达水平分别为0.37±0.11,0.82±0.13和0.79±0.12,FOXL2蛋白相对表达水平分别为0.53±0.08,0.90±0.1和0.87±0.13,ITGA2蛋白相对表达水平分别为0.45±0.07,0.93±0.14和0.91±0.15。实验组的上述指标与空白组和对照组比较,差异均有统计学意义(均P<0.05)。结论过表达miR-329可靶向下调E2F1基因表达,抑制非小细胞肺癌A549细胞顺铂耐药性,其可能是通过抑制FOXL2/ITGA2通路活化实现的。Objective To investigate the effect of miR-329 targeting cell cycle related transcription factor(E2F1)gene on cisplatin(DDP)resistance in non-small cell lung cancer A549 cells.Methods The DDP resistant A549/DDP cells were divided into blank group(without any treatment),control group(transfected with miR-329-mimics-NC)and experimental group(transfected with miR-329-mimics).The expression levels of miR-329 and E2F1 mRNA were measured by real-time quantitative polymerase chain reaction.Cell proliferation was detected by cell counting kit-8.E2F1 protein and recombinant human forkhead box protein L2/integrinα2(FOXL2/ITGA2)pathway protein expression were detected by Western blot.Results The relative expression levels of miR-329 in A549 cells and A549/DDP cells were 1.00±0.16 and 0.53±0.08 with significant difference(P<0.05).The relative expression levels of miR-329 in the experimental,control and blank groups were 1.68±0.26,1.02±0.16 and 1.00±0.15;the relative expression levels of E2F1 mRNA were 0.33±0.05,1.01±0.15 and 1.00±0.16;the cell proliferation inhibition rates were(28.95±4.33)%,(14.03±2.10)%and(13.26±1.98)%;the half-inhibitory concentrations were(28.40±1.72),(47.85±5.71)and(45.63±3.18)μmol·L^(-1);the relative expression levels of E2F1 protein were 0.37±0.11,0.82±0.13 and 0.79±0.12;the relative expression levels of FOXL2 protein were 0.53±0.08,0.90±0.13 and 0.87±0.13;the relative expression levels of TGA2 protein were 0.45±0.07,0.93±0.14 and 0.91±0.15,respectively.Compared with the blank and control groups,the above indexes of experimental group had significant differences(all P<0.05).Conclusion Overexpression of miR-329 can target down regulate E2F1 gene expression and inhibit cisplatin resistance in nonsmall cell lung cancer A549 cells,which may be achieved by inhibiting the activation of FOXL2/ITGA2 pathway.

关 键 词：顺铂耐药 非小细胞肺癌 微小核糖核苷酸-329 细胞周期相关转录因子 

分 类 号：R979.1[医药卫生—药品]