circ_0061140靶向miR-6838-5p促进非小细胞肺癌细胞的增殖、阻滞细胞周期、诱导细胞凋亡  

circ_0061140 Regulate Proliferation,Cell Cycle and Apoptosis of Non-small Cell Lung Cancer Cells by Targeting miR-6838-5p

作　　者：蒋君霞 杨国荣[2] 谷涉群[2] 喻珣 JIANG Junxia;YANG Guorong;GU Shequn;YU Pei(First Ward,Department of Oncology,Chenzhou First People’s Hospital,Chenzhou,Hunan,423000,China;Department of Oncology,Chenzhou First People’s Hospital,Chenzhou,Hunan,423000,China)

机构地区：[1]郴州市第一人民医院肿瘤内科一区,湖南省郴州市423000 [2]郴州市第一人民医院肿瘤内科,湖南省郴州市423000

出　　处：《医学分子生物学杂志》2022年第6期464-471,共8页Journal of Medical Molecular Biology

摘　　要：目的通过实验研究确认环状RNA(circRNA)circ_0061140是否能靶向miR-6838-5p调控非小细胞肺癌细胞的增殖、细胞周期和凋亡。方法采用逆转录-定量聚合酶链反应(RT-qPCR)检测circ_0061140和miR-6838-5p在非小细胞肺癌细胞中的表达情况。将非小细胞肺癌NCI-H1299细胞分为si-circ_0061140组(转染si-circ_0061140;低表达circ_0061140)、si-NC组(转染si-NC;阴性对照)、NC组(未转染;空白对照)、miR-6838-5p组(转染miR-6838-5p模拟物;高表达miR-6838-5p)、miR-NC组(转染miR-NC;高表达miR-6838-5p的阴性对照)、si-circ_0061140+anti-miR-NC组(共转染si-circ_0061140与anti-miR-NC)、si-circ_0061140+anti-miR-6838-5p组(共转染si-circ_0061140与anti-miR-6838-5p)。采用Western印迹检测细胞周期蛋白D1(cyclinD1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶-3(cleavedcaspase-3)蛋白表达,MTT法检测细胞增殖能力,流式细胞仪检测细胞周期和凋亡情况。双荧光素酶活性检测circ_0061140和miR-6838-5p的靶向结合。结果非小细胞肺癌组织、细胞NCI-H1299、NCI-H2170、NCI-H1975中的circ_0061140表达量均比癌旁组织或正常肺上皮细胞BEAS-2B高,miR-6838-5p表达量比癌旁组织或BEAS-2B细胞低(P_(均)<0.05)。si-circ_0061140组或miR-6838-5p组NCI-H1299细胞的CyclinD1蛋白表达量、增殖能力、S期细胞比例比NC组减少,Cleaved-caspase-3蛋白表达量、G_(0)/G_(1)期细胞比例、凋亡率比si-NC组或miR-NC组增加(P均<0.05)。circ_0061140靶向调控miR-6838-5p的表达。共转染si-circ_0061140、anti-miR-6838-5p可减弱转染si-circ_0061140对细胞生物学行为的作用。结论低表达circ_0061140通过靶向非小细胞肺癌细胞的miR-6838-5p,抑制细胞增殖、阻滞G_(0)/G_(1)期细胞周期,并加速凋亡。Objective To investigate the effect of circular RNA(circRNA)circ_0061140 on the proliferation,cell cycle and apoptosis of non-small cell lung cancer cells via targeting miR-6838-5p.Methods Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of circ_0061140 and miR-6838-5p in the non-small cell lung cancer cells.The NCI-H1299 non-small cell lung cancer cells were divided into 7 groups:the si-circ_0061140 group(transfected with si-circ_0061140,circ_0061140 lowly expressed),the si-NC group(transfected with si-NC,the negative control),the NC group(blank control with no transfection of miRNA),the miR-6838-5p group(transfected with miR-6838-5p mimic,miR-6838-5p highly expressed),the miR-NC group(transfected with miR-NC,the negative control),the sicirc_0061140+anti-miR-NC group(co-transfected with si-circ_0061140 and anti-miR-NC),the si-circ_0061140+anti-miR-6838-5p group(co-transfected with si-circ_0061140 and anti-miR6838-5p).Western blotting was used to detect the expression levels of cyclin D1 and cleaved cysteinyl aspartate specific proteinase-3(cleaved-caspase-3).The proliferation of cells was detected by MTT method.The cell cycle and cell apoptosis was detected by the flow cytometry.Dual luciferase reporter assay was used to verify the relationship between circ_0061140 and miR-6838-5p.Results The expression level of circ_0061140 in the non-small cell lung cancer tissues and the cell lines NCI-H1299,NCI-H2170,NCI-H1975 were all higher than that in the adjacent tissues and the BEAS-2B normal lung epithelial cells,while the expression level of miR-6838-5p in the non-small cell lung cancer tissues and the lung cancer cells was lower than that in the adjacent tissues and BEAS2B cells(all P<0.05).the expression level of cyclin D1 protein,the proliferation rate,and the ratio of S phase cells in cells of the si-circ_0061140 group and the miR-6838-5p group were lower than those of the NC group,while the expression level of cleaved-caspase-3 protein,the ratio of G_(

关 键 词：circ_0061140 非小细胞肺癌 miR-6838-5p 细胞增殖 细胞周期 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]