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.
作者
高昕
杨智
于海波
才陆贤
王知勇
刘洪汐
王利斌
刘博
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)
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)~~
