摘要
目的探讨微小RNA(miRNA, miR)-597靶向Rab23对肝癌细胞增殖、凋亡和周期的影响及其分子机制。方法选取南阳市中心医院2017年5月到2019年5月收集的126例肝癌和癌旁组织作为研究对象, 采用荧光定量聚合酶链反应(PCR)分析正常肝细胞LO2、肝癌细胞系HepG2、SMMC-7721和Huh7中miR-597表达水平;采用慢病毒在肝癌细胞系HepG2构建对照miRNA和miR-597过表达细胞系, 分别为对照组和观察组, 采用细胞计数试剂盒(CCK-8)、克隆形成实验和异体移植瘤实验分析肿瘤细胞的增殖能力;采用流式细胞术分析细胞周期和凋亡水平;采用生物信息学和双荧光素酶报告基因分析miR-597的靶基因;采用蛋白质印迹法(Western blot)检测靶蛋白表达水平。组间计量资料比较采用t检验。结果肝细胞LO2中miR-597表达水平(1.09±0.15)明显高于肝癌细胞HepG2、SMMC-7721和Huh7 (0.37±0.14、0.45±0.16、0.58±0.14), 差异有统计学意义(t=4.011、3.714、3.281, P<0.05)。对照组细胞吸光度值(2.87±0.26)明显高于观察组(1.53±0.22), 差异有统计学意义(t=3.091, P<0.05)。对照组细胞克隆形成率[(71.23±8.29)%]明显高于观察组[(38.54±4.98)%], 差异有统计学意义(t=5.220, P<0.05)。对照组小鼠体内肿瘤体积[(1 284.59±209.32) mm^(3)]明显高于观察组[(799.58±109.43) mm^(3)], 差异有统计学意义(t=5.530, P<0.05)。对照组小鼠肿瘤重量[(3.98±0.29) g]明显高于观察组[(1.42±0.19) g], 差异有统计学意义(t=3.218, P<0.05)。对照组细胞S期比例[(34.21±4.58)%]明显高于观察组[(44.58±5.87)%], 差异有统计学意义(t=4.719, P<0.05)。对照组细胞G2期比例[(32.14±4.28)%]低于观察组[(20.37±4.29)%], 差异有统计学意义(t=3.612, P<0.05)。对照组细胞凋亡率[(3.76±1.09)%]明显低于观察组[(24.54±3.68)%], 差异有统计学意义(t=4.006, P<0.05)。生物信息学和双荧光素酶报告基因结果显示, Rab23是miR-597的靶基因。对照组细胞Rab23蛋白表达水�
Objective To investigate the effects of Rab23 targeted by microRNA(miRNA,miR)-597 on proliferation,apoptosis and cycle of hepatoma cells and its molecular mechanism.Methods A total of 126 cases of liver cancer and para-cancerous tissues collected in Nanyang Central Hospital from May 2017 to May 2019 were selected as the research objects.The expression levels of miR-597 in normal hepatic cells LO2,and hepatoma cell lines HepG2,SMMC-7721 and Huh7 were analyzed by fluorescence quantitative polymerase chain reaction(PCR).Lentivirus was used to construct control miRNA and miR-597 overexpression cell lines in hepatoma cell line HepG2,serving as control group and observation group respectively.The proliferation ability of tumor cells was analyzed by cell counting kit-8(CCK-8),clone formation experiment and allogeneic transplantation experiment.The cell cycle and apoptosis were analyzed by flow cytometry.The target gene of miR-597 was analyzed by bioinformatics and double luciferase reporter gene.The expression level of target protein was detected by Western blotting.The measurement data between groups were compared by t-test.Results The expression level of miR-597 in LO2(1.09±0.15)was significantly higher than that in HepG2,SMMC-7721 and Huh7(0.37±0.14;0.45±0.16;0.58±0.14,t=4.011,3.714,3.281,P<0.05).The light absorption value of cells in the control group(2.87±0.26)was significantly higher than that in the observation group(1.53±0.22,t=3.091,P<0.05).The clone formation rate of cells in the control group[(71.23±8.29)%]was significantly higher than that in the observation group[(38.54±4.98)%,t=5.220,P<0.05].The tumor volume in the control group[(1284.59±209.32)mm3]was significantly greater than that in the observation group[(799.58±109.43)mm3,t=5.530,P<0.05].The tumor weight of the control group[(3.98±0.29)g]was significantly greater than that of the observation group[(1.42±0.19)g,t=3.218,P<0.05].The proportion of S-phase cells in the control group[(34.21±4.58)%]was significantly higher than that in the obser
作者
梅孝臣
王娟
王磊
厉冰
门中俊
侯潇峰
陈磊
孙万日
Mei Xiaochen;Wang Juan;Wang Lei;Li Bing;Men Zhongjun;Hou Xiaofeng;Chen Lei;Sun Wanri(Department of General Surgery,Nanyang Central Hospital/Department of General Surgery,Nanyang Central Hospital Affiliated to Zhengzhou University,Nanyang 473000,China;Department of General Surgery,First People’s Hospital,Nanyang 473000,China;Department of Pediatrics,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第2期265-268,共4页
Chinese Journal of Experimental Surgery
关键词
微小RNA
肝癌
增殖
周期
凋亡
MicroRNA
Liver cancer
Proliferation
Cycle
Apoptosis
