shRNA-p53对p125表达及肝癌SMMC-7721细胞增殖的影响  

Effect of shRNA-p53 on p125 expression and proliferation of SMMC-7721 cells

作　　者：廖柳凤[1] 欧贤红[2] 吴琼[1] 徐恒 李艳[1] 刘华钢[2] LIAO Liu-feng;OU Xian-hong;WU Qiong;XU Heng;LI Yan;LIU Hua-gang(Department of Pharmacy,Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Department of Pharmacy,Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;School of Medicine,Emory University,Atlanta 30328,USA)

机构地区：[1]广西医科大学附属肿瘤医院药学部,广西壮族自治区南宁530021 [2]广西医科大学药学院,广西壮族自治区南宁530021 [3]School of Medicine,Emory University,Atlanta 30328,USA

出　　处：《中国临床药理学杂志》2020年第16期2428-2431,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81260657)。

摘　　要：目的研究p53对p125表达的影响,及其对人肝癌细胞SMMC-7721细胞增殖的影响。方法用脂质体转染技术,将实验分为3组:对照组(转染空载体pGPU6/GFP/neo-shNC的7721-shNC细胞)、空白组(肝癌细胞SMMC-7721)、实验组(转染p53重组真核表达质粒pGPU6/GFP/neo-p53-shRNA的7721-p53-shRNA细胞)。用蛋白免疫印迹法(Western Blotting)检测p53和p125蛋白表达水平,用实时荧光聚合酶链反应检测DNA聚合酶δ催化亚基基因1(POLD1)、视网膜母细胞瘤蛋白1(Rb1)、转录因子E2F、细胞周期依赖性蛋白激酶2(CDK2)及细胞周期素E(CylinE)基因的表达水平。结果实验组、对照组和空白组的p53蛋白表达水平分别为0.22±0.03,0.55±0.07和0.53±0.06,p125蛋白分别为2.86±0.21,2.20±0.28和2.24±0.25,POLD1 mRNA分别为1.09±0.13,0.45±0.07和1.00±0,Rb1 mRNA分别为4.16±0.29,1.10±0.16和1.00±0,转录因子E2F mRNA分别为2.49±0.12,1.08±0.17和1.00±0,CDK2 mRNA分别为1.65±0.08,1.12±0.22和1.00±0,CyclinE mRNA分别为2.78±0.23,1.17±0.31和1.00±0,实验组的上述指标与对照组比较,差异均有统计学意义(均P<0.05)。结论通过RNA干扰技术反向验证了p53对p125表达的调控作用,其对肝癌细胞增殖具有抑制作用,可能是通过调控Rb1、转录因子E2F、CDK2和CyclinE等细胞周期因子基因表达而实现的。Objective To study the effects of p53 on the expression of p125 and the proliferation of SMMC-7721 cells. Methods By using liposome transfection technology, the experiment was divided into three groups: control group(7721-shNC cells transfected with empty vector pGPU6/GFP/neo-shNC), blank group(SMMC-7721 cells) and experimental group(7721-p53-shRNA cells transfected with p53 eukaryotic expression plasmid pGPU6/GFP/neo-p53-shRNA). The expression levels of p53 and p125 protein were detected by Western Blotting. The expressions of DNA polymerase δ catalytic subunit 1(POLD1), retinoblastoma protein 1(Rb1), transcription factor E2F, cell cycle dependent protein kinase 2(CDK2) and CyclinE mRNA were detected by real time fluorescence polymerase chain reaction. Results The expression levels of p53 protein in experimental, control and blank groups were 0.22±0.03, 0.55±0.07 and 0.53±0.06, p125 protein were 2.86±0.21, 2.20±0.28 and 2.24±0.25, POLD1 mRNA were 1. 09 ± 0. 13,0. 45 ± 0. 07 and 1. 00 ± 0,Rb1 mRNA were 4. 16 ± 0. 29,1. 10 ± 0. 16 and 1. 00 ± 0,transcription factor E2F mRNA were 2. 49 ± 0. 12,1. 08 ± 0. 17 and 1. 00 ± 0,CDK2 mRNA were 1. 65 ± 0. 08,1. 12 ± 0. 22 and1. 00 ± 0,CyclinE mRNA were 2. 78 ± 0. 23,1. 17 ± 0. 31 and 1. 00 ± 0. Compared with the control group,the above indexes in the experimental group had statistically significant differences(all P < 0. 05). Conclusion The regulatory effect of p53 on p125 expression and its inhibitory effect on the proliferation of hepatoma cells were reverse verified by RNA interference technique,which may be realized by regulating the expression of cell cycle factor genes such as Rb1,transcription factor E2F,CDK2 and CyclinE.

关 键 词：DNA聚合酶Δ P53 p125 肝癌细胞 

分 类 号：R97[医药卫生—药品]