微小核糖核酸let-7c对肝癌细胞增殖的影响及其靶基因分析  

Effect of micro RNA let-7c on proliferation of hepatocellular carcinoma cells and analysis of its target genes

作　　者：祖珊珊 Zu Shanshan(Department of Clinical Laboratory,Longnan Hospital of Daqing City,Daqing,Heilongjiang 163453,China)

机构地区：[1]大庆龙南医院检验科,黑龙江省163453

出　　处：《中国基层医药》2020年第12期1418-1421,共4页Chinese Journal of Primary Medicine and Pharmacy

摘　　要：目的分析微小核糖核酸(mi RNA)let-7c通过靶定细胞周期蛋白依赖激酶6(CDK6)抑制肝癌细胞增殖的临床意义,并探讨其对肝癌细胞增殖的影响以及靶基因。方法选取人低转移肝癌细胞-97L(MHCC-97L)、人高转移肝癌细胞(HCCLM3)、人肝癌组织(HepG2)、人肝癌细胞-7721(SMMC-7721)、人肝细胞LO2、小鼠抗人细胞周期蛋白依赖激酶抗体等进行细胞培养,利用Western blot检测转染后小鼠CDK6表达情况,转染48 h后应用细胞裂解液对三组蛋白进行提取,将材料分为观察组、对照组、空白组,观察组转染let-7c,对照组转染阴性对照miRNA,空白组未进行转染处理;应用二喹啉甲酸法对各孔细胞蛋白浓度进行测定。结果let-7c对肝癌细胞HCCLM3增殖结果显示,于450 nm处观察组、对照组转染后24 h吸光度值差异无统计学意义(P>0.05);转染48 h后观察组、对照组HCCLM3细胞中CDK6蛋白含量发生较大变化,观察组的CDK6蛋白表达量为(0.58±0.05),均低于对照组的(0.81±0.08)与空白组(0.85±0.09)(t=10.107、13.876,均P<0.05);转染后72 h,观察组细胞处于G1期比例为(56.21±3.25)%,明显高于对照组的(45.21±1.56)%与空白组的(46.24±1.98)%,差异均有统计学意义(t=16.146、13.862,均P=0.000)。结论肝癌细胞中let-7c呈低表达,将let-7c上调可有效调控CDK6,从而抑制癌细胞生长,对肝癌细胞增殖具有明显的抑制作用。Objective To analyze the clinical significance of let-7c microribonucleic acid(miRNA)inhibiting the proliferation of hepatocellular carcinoma cells by targeting cyclin-dependent kinase 6(CDK6),and to explore its effect on the proliferation of hepatocellular carcinoma cells and target genes.Methods Human low metastatic hepatocellular carcinoma cell line-97L(MHCC-97L),human high metastatic hepatocellular carcinoma cell line(HCCLM3),human hepatocellular carcinoma tissue(HepG2),human hepatocellular carcinoma cell line-7721(SMMC-7721),human hepatocyte LO2 and mouse anti-human cyclin dependent kinase antibody were selected for cell culture.The expression of CDK6 in transfected mice was detected by Western blot.After 48 hours of transfection,three histones were extracted by cell lysate.The materials were divided into observation group,control group and blank group.The observation group was transfected with let-7c,while the control group was transfected with negative control microRNAs and the blank group was not transfected.The protein concentration of porous cells was determined by diquinoline formic acid method.Results The results of let-7c on HCCLM3 proliferation showed that there was no statistically significant difference in the absorbance value at 450nm between the observation group and the control group 24 hours after transfection(P>0.05).After 48 hours of transfection,the content of CDK6 protein in the observation group and the control group changed greatly,and the expression of CDK6 protein in the observation group(0.58±0.05)was lower than that in the control group(0.81±0.08)and blank group(0.85±0.09)(t=10.107,13.876,all P<0.05).At 72 hours after transfection,the percentage of cells in G1 phase in the observation group[(56.21±3.25)%]was significantly higher than that in the control group[(45.21±1.56)%]and blank group[(46.24±1.98)%],the differences were statistically significant(t=16.146,13.862,all P=0.000).Conclusion The expression of let-7c in hepatocellular carcinoma cells is low.Upregulation of let-7c c

关 键 词：肝肿瘤 核糖核酸酶类 基因 Let-7c 细胞周期蛋白依赖激酶6 细胞增殖 肿瘤转移 逆转录病毒科 基因 肿瘤抑制 肿瘤细胞 培养的 

分 类 号：R735.7[医药卫生—肿瘤]