摘要
目的探讨环状RNA(CircRNA)-1565通过10-11转位基因2(TET2)基因对非小细胞肺癌A549细胞增殖、细胞周期和凋亡的影响。方法采用qPCR检测CircRNA-1565在肺癌组织、癌旁组织、不同肺癌细胞系中的表达,同时选取CircRNA-1565高表达的A549细胞系,并分为对照组(正常A549细胞)、阴性对照组(转染无基因序列的A549细胞)、实验组(转染沉默CircRNA-1565质粒的A549细胞)。采用MTT检测各组细胞增殖活性,流式细胞术检测细胞周期、凋亡率,Western blot检测各组细胞TET2、细胞周期蛋白D1/E1(Cyclin D1/E1)、细胞周期素依赖性激酶1/2(CDK1/2)、凋亡蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、多聚ADP核糖聚合酶(PARP)的表达情况。结果与癌旁组织比较,肺癌组织CircRNA-1565表达明显上调(P<0.05);与对照组和阴性对照组比较,实验组A549细胞增殖活性明显降低,G_(0)/G_(1)、G_(2)/M期细胞数量明显增加,S期细胞明显减少,细胞凋亡率显著升高(P<0.05);Western blot结果显示实验组细胞TET2蛋白、细胞周期蛋白Cyclin D1/E1、CDK1/2表达水平明显下调,凋亡蛋白Bax、PARP表达水平明显上调(P<0.05)。结论CircRNA-1565可促进非小细胞肺癌A549细胞增殖、抑制细胞凋亡,机制可能与下调TET2基因的表达、调控细胞周期蛋白、激活凋亡蛋白通路的表达有关。
Objective To explore the effect of CircRNA-1565 on the proliferation,cell cycle and apoptosis of non-small cell lung cancer A549 through ten-eleven translocation 2(TET2)gene.Methods QPCR was used to detect the expression of CircRNA-1565 in lung cancer tissues,adjacent tissues,and different lung cancer cell lines.At the same time,the A549 cell line with high expression of CircRNA-1565 was selected and divided into the control group(normal A549 cells),the negative control group(transition)A549 cells transfected with negative plasmid and the experiment group(transfected with silencing CircRNA-1565 plasmid A549 cells).MTT was used to detect cell proliferation activity in each group,cell cycle and apoptosis rate were detected by flow cytometry,and Western blot was used to detect the expression of TET2,cyclin D1/E1(Cyclin D1/E1),cyclin-dependent kinase 1/2(CDK1/2),B-cell lymphoma 2(Bcl-2),Bcl-2 related X protein(Bax),poly ADP-ribose polymerase(PARP).Results Compared with the adjacent tissues,the expression of CircRNA-1565 in lung cancer tissue was significantly up-regulated(P<0.05);compared with the control group and the negative control group,the proliferation activity of A549 cells in the experiment group significantly reduced,the number of cells in the G_(0)/G_(1),G_(2)/M phase significantly increased,S phase cells significantly reduced,and the apoptosis rate significantly increased(P<0.05);Western blot results showed that the expression levels of TET2 protein,Cyclin D1/E1,and CDK1/2 in the experiment group were significantly down-regulated,and the expression level of the apoptotic protein Bax,PARP significantly increased(P<0.05).Conclusion CircRNA-1565 can promote the proliferation and inhibit apoptosis of non-small cell lung cancer A549 cells.The mechanism may be related to down regulating the expression of TET2 gene,regulating cell cycle proteins and activating the expression of apoptotic protein pathway.
作者
张茹茹
朱正秋
彭猛青
徐娟
张坤
ZHANG Ruru;ZHU Zhengqiu;PENG Mengqing;XU Juan;Zhang Kun(Department of Oncology,Suqian Zhongwu Hospital/Suqian Tumor Hospital,Suqian,Jiangsu 223800,China;Department of Oncology,Affiliated Hospital of Xuzhou Medical University,Xuzhou,Jiangsu 221000,China)
出处
《重庆医学》
CAS
2021年第24期4156-4161,共6页
Chongqing medicine
基金
江苏省宿迁市指导性科技计划项目(Z2020042)。
