微小RNA-504对非小细胞肺癌细胞增殖及凋亡的作用  

Effect of microRNA-504 on proliferation and apoptosis of non-small cell lung cancer cells

作　　者：高昕 杨智 于海波 才陆贤 王知勇 刘洪汐 王利斌 刘博[2] GAO Xin1, YANG Zhi1, YU Haibo1, CAI Luxian1, WANG Zhiyong1, LIU Hongxi1, WANG Libin1, LIU Bo2(1. Department of Thoracic Surgery, Northeast International Hospital, Shenyang 110623; 2. Department of Thoracic Surgery, General Hospital of Shenyang Military Region, Shenyang 110016, Chin)

机构地区：[1]东北国际医院胸外科,沈阳110623 [2]沈阳军区总医院胸外科,沈阳110016

出　　处：《临床与病理杂志》2018年第2期239-245,共7页Journal of Clinical and Pathological Research

基　　金：辽宁省科学技术计划项目(2012225019)~~

摘　　要：目的:探讨微小RNA-504(microRNA-504,miR-504)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖及凋亡的影响及其机制。方法:以定量PCR检测3种NSCLC细胞株A549,H1299,HCC827及对照人支气管上皮细胞株BEAS-2B中miR-504的表达。选取miR-504变化最明显的细胞株(HCC827)用于后续实验。用Lipofectamine RNAiMAX分别将miR-504类似物(miR-504mimic)、miR-504抑制物(miR-504 inhibitor)及非靶向miRNA对照(miR-control)转染至HCC827细胞中。以CCK-8试剂盒检测上述3组细胞增殖情况。用Annexin V-FITC/PI凋亡试剂盒检测3组细胞凋亡情况。以Western印迹法检测3组细胞P53表达。结果:与对照组BEAS-2B细胞相比,miR-504在3种NSCLC细胞株中表达水平均显著增加,其中以HCC827细胞增加幅度最大。与miR-control组相比,过表达miR-504(miR-504 mimic)后HCC827细胞增殖加快、凋亡减少,而抑制miR-504表达(miR-504 inhibitor)后HCC827增殖减慢、凋亡增加。Western印迹检测结果显示:与miR-control组相比,P53在miR-504 mimic组表达水平降低,而在miR-504 inhibitor组表达水平增加。结论:miR-504具有促进NSCLC细胞增殖、抑制凋亡的作用,其机制可能与下调P53表达有关。Objective： To investigate the effect and possible mechanism of microRNA-504（miR-504） in proliferation and apoptosis of non-small cell lung cancer（NSCLC） cells. Methods： Expression of miR-504 in three NSCLC cell lines（A549, H1299 and HCC827） and control human bronchial epithelial cell line BEAS-2 B were determined by real-time PCR. The cell line（HCC827） with the most significant change of miR-504 expression was used in the following study. Then HCC827 cells were transfected with miR-504 mimic, miR-504 inhibitor and nontargeting miR-control by using Lipofectamine RNAi MAX. Cell proliferation of the three above-mentioned groups was detected by CCK-8 kit. Cell apoptosis of the three groups was measured by Annexin V-FITC/PI apoptosis detection kit. Expression of P53 of the three groups was determined by Western blot. Results： Compared with BEAS-2 B control cells, expression of miR-504 was significantly increased in all three NSCLC cell lines. HCC827 cells were used in the following study due to its most significant increase of miR-504. Compared with miR-control group, over-expression of miR-504（miR-504 mimic） resulted in accelerated proliferation and decreased apoptosis of HCC827 cells. While down-regulation of miR-504（miR-504 inhibitor） led to retarded proliferation and increased apoptosis. In addition, expression of P53 was decreased in miR-504 mimic group but increased in miR-504 inhibitor group. Conclusion： miR-504 could promote proliferation and inhibit apoptosis of NSCLC cells, which was possibly mediated by down-regulating P53 expression.

关 键 词：微小RNA-504 非小细胞肺癌 增殖 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]