微小RNA-342-3p靶向性别决定区Y框蛋白6对肝癌细胞增殖、周期和凋亡的影响  

The effect of microRNA-342-3p targeting sex determining region Y-box 6 on the proliferation, cycle, and apoptosis of liver cancer cells

作　　者：王艳瑛 吴伟[1] 王亚奇 高志强[2] 马鹏 郭凯[1] Wang Yanying;Wu Wei;Wang Yaqi;Gao Zhiqiang;Ma Peng;Guo Kai(Department of Hepatobiliary,Pancreatic and Splenic Surgery,Zhumadian Central Hospital Affiliated to Huanghuai University,Zhumadian 463000,China;Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]黄淮学院附属驻马店市中心医院肝胆胰脾外科,驻马店463000 [2]河南省郑州大学第一附属医院肝胆胰外科,郑州450052

出　　处：《中华实验外科杂志》2023年第11期2237-2240,共4页Chinese Journal of Experimental Surgery

基　　金：2021年河南省高等学校重点科研项目(21A320061)。

摘　　要：目的:探讨微小RNA(miRNA,miR)-342-3p靶向性别决定区Y框蛋白6(SOX6)对肝癌细胞增殖、周期和凋亡的影响及其分子机制。方法:选取2021年6月至2022年6月驻马店市中心医院收集的121例原发性肝癌组织和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)技术分析肝癌组织和癌旁组织miR-342-3p表达水平;采用miRNA对照和miR-342-3p过表达慢病毒感染人肝癌细胞HepG2,构建miRNA对照组和miR-342-3p组细胞,采用细胞计数试剂盒(CCK-8)、克隆形成实验和体外移植瘤实验分析miRNA对照组和miR-342-3p组细胞的增殖;采用流式细胞术分析两组细胞的周期变化;采用膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)/碘化丙锭(PI)凋亡试剂盒分析两组细胞的凋亡水平;采用生物信息学和双荧光素酶报告基因分析miR-342-3p的靶基因;蛋白质免疫印迹(Western blot)分析蛋白质表达水平。组间计数资料比较采用t检验。结果:肝癌组织中miR-342-3p表达水平(1.23±0.22)明显低于癌旁组织表达水平(0.71±0.14),差异有统计学意义(t=21.710,P<0.05)。miRNA对照组细胞吸光度(A)值(2.02±0.13)明显高于miR-342-3p组(1.61±0.09),差异有统计学意义(t=6.441,P<0.05)。miRNA对照组细胞克隆形成率[(85.92±4.95)%]明显高于miR-342-3p组[(59.06±4.39)%],差异有统计学意义(t=9.942,P<0.05)。miRNA对照组细胞体内成瘤体积[(690.67±72.49)mm 3]明显高于miR-342-3p组[(59.06±4.39)mm 3],差异有统计学意义(t=9.942,P<0.05)。miRNA对照组细胞G 0/G 1期百分比[(41.33±2.58)%]明显低于miR-342-3p组[(58.33±2.93)%],差异有统计学意义(t=10.980,P<0.05)。miRNA对照组细胞S期百分比[(39.17±2.71)%]明显高于miR-342-3p组[(28.33±3.31)%],差异有统计学意义(t=5.911,P<0.05)。miRNA对照组细胞凋亡率[(2.63±0.95)%]明显低于miR-342-3p组[(16.33±2.29)%],差异有统计学意义(t=13.510,P<0.05)。SOX6是miR-342-3p的靶基因。癌旁组织中miR-342-3p靶基因SOX6表达水平(1.01±Objective To investigate the effects of microRNA(miRNA,miR)-342-3p targeting sex determining region Y-box 6(SOX6)on the proliferation,cycle,and apoptosis of liver cancer cells and its molecular mechanisms.Methods 121 primary liver cancer tissues and adjacent tissues collected from Zhumadian Central Hospital from June 2021 to June 2022 were selected as the research subjects.The expression level of miR-342-3p in liver cancer tissues and adjacent tissues were analyzed by fluorescence quantitative polymerase chain reaction(PCR)technology.Using miRNA control and miR-342-3p overexpression lentivirus to infect human liver cancer cell line HepG2,miRNA control group and miR-342-3p group cells were constructed.The proliferation of miRNA control group and miR-342-3p group cells was analyzed by cell counting kit 8(CCK-8),clone formation assay,and in vitro tumor transplantation assay.The cycle changes of the two groups of cells was analyzed by flow cytometry.The apoptosis levels of the two groups of cells were analyzed using Annexin V-fluoresceine isothiocyanate(FITC)/propidium iodide(PI)apoptosis kit.Analyze the target genes of miR-342-3p using bioinformatics and dual luciferase reporter genes;Western blotting analysis of protein expression levels.The comparison of inter group counting data was conducted using t-test.Results The expression level of miR-342-3p in liver cancer tissue(1.23±0.22)was significantly lower than that in adjacent tissues(0.71±0.14,t=21.710,P<0.05).The absorbance(A)value of cells in the miRNA control group(2.02±0.13)was significantly higher than that in the miR-342-3p group(1.61±0.09,t=6.441,P<0.05).The cell clone formation rate in the miRNA control group[(85.92±4.95)%]was significantly higher than that in the miR-342-3p group[(59.06±4.39)%,t=9.942,P<0.05].The tumor formation volume in the miRNA control group cells[(690.67±72.49)mm3]was significantly higher than that in the miR-342-3p group cells[(59.06±4.39)mm3,t=9.942,P<0.05].The percentage of G0/G1 phase cells in the miRNA control group[(41.

关 键 词：微小RNA-342-3p 性别决定区Y框蛋白6 肝癌 增殖 周期 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]