干预Wip1基因对人肝癌细胞系Huh-7的影响  

Influence of intervention of Wip1 gene on biological characteristics of human hepatoma cell line Huh-7

作　　者：李哲[1] 刘军[2] 李晓明[1] 赵丽娟[1] 路健[1] 吴灵潼[1] 李秉寿[1] 

机构地区：[1]山东大学附属省立医院普外科 [2]山东大学附属省立医院器官移植肝胆外二科

出　　处：《中国临床研究》2016年第10期1297-1302,共6页Chinese Journal of Clinical Research

基　　金：国家自然基金面上项目(81373172)

摘　　要：目的干预人肝癌细胞系Huh-7的野生型p53诱导的磷酸酶1（Wip1）基因的表达,并观察其细胞生物学特性的变化。方法采用转染法,将促进Wip1基因表达的质粒和抑制Wip1基因表达的小干扰RNA（siRNA）分别转染Huh-7细胞。质粒转染的Huh-7细胞为质粒转染组,未经质粒转染的为未转染组,siRNA转染的为siRNA干扰组,用Negative Control siRNA转染的为NC组。转染后,行Western blot、CCK-8、细胞划痕、Transwell实验,检测各组细胞生物学特性的变化。结果质粒转染后,Huh-7细胞Wip1蛋白的表达上调,细胞增殖促进,增殖率在107.42%-176.36%之间。未转染组在0-24 h、-48 h、-72 h时的迁移距离分别小于质粒转染组：[（34.78±4.77）μm vs（61.60±2.02）μm,（33.98±3.48）μm vs（64.75±4.67）μm,（35.49±0.90）μm vs（67.89±4.23）μm,P均〈0.01]。未转染组3次Transwell实验的细胞迁移数均少于质粒转染组：[（90.00±3.00）vs（178.67±5.77）,（74.33±26.95）vs（209.00±32.91）,（79.33±20.74）vs（208.67±73.24）,P均〈0.01]。siRNA转染后,Huh-7细胞Wip1蛋白的表达下调;细胞抑制率在12.37%-40.81%之间;NC组在0-24 h、-48 h、-72 h时的迁移距离分别大于siRNA干扰组：[（31.24±4.42）μm vs（16.04±6.21）μm,（31.33±2.13）μm vs（18.20±5.67）μm,（32.45±3.08）μm vs（18.99±5.32）μm,P均〈0.01];NC组3次Transwell实验的细胞迁移数均多于siRNA干扰组：[（74.00±12.17）vs（33.00±10.44）,（77.00±5.00）vs（32.67±2.08）,（81.33±17.16）vs（29.30±2.51）,P均〈0.01]。结论促进Huh-7细胞Wip1基因表达后,其Wip1蛋白的表达上调,细胞增殖、迁移、侵袭能力增强。抑制Huh-7细胞Wip1基因表达后,其Wip1蛋白的表达下调,细胞增殖、迁移、侵袭能力被抑制。Objective To observe the changes of biological characteristics of human hepatoma cell line Huh-7 by intervening the expession of its wild type p53 induced-phosphatases 1（ Wipl） gene. Methods Using transfection method,the plasmid promoting Wip1 gene expression and small interfering ribonucleic acid（ siRNA） inhibiting Wip1 gene expression were respectively transfected into Huh-7cells. The Huh-7 cells with plasmid transfection were served as plasmid transfection group; the Huh-7 cells without plasmid transfection were served as non-transfection group; the Huh-7cells with siRNA transfection were served as siRNA interference group,the Huh-7cells with negative control siRNA transfection were served as NC group. After transfection,Western blot test,cell counting kit-8（ CCK-8）,cell wound scratch assay and Transwell experiment were used to detect the biological characteristics of Huh-7 cells. Results After plasmid transfection,the Wip1 protein expression was up-regulated,and the cells proliferation was enhanced（ cell proliferation rate was between 107. 42%-176.36%）. The cell migration distances at 0- 24 h,24- 48 h,48- 72 h after transfection in non-transfection group were less than those in plasmid transfection group,respectively [（ 34. 78 ± 4. 77） μm vs（ 61. 60 ± 2. 02） μm,（ 33. 98 ± 3. 48） μm vs（ 64. 75 ± 4. 67） μm,（ 35. 49 ± 0. 90） μm vs（ 67. 89 ± 4. 23） μm,all P〈0. 01]. The cell migration numbers in the Transwell experiments of 3 times in non-transfection group were all less than those in plasmid transfection group [（ 90. 00 ±3. 00） vs（ 178. 67 ± 5. 77）,（ 74. 33 ± 26. 95） vs（ 209. 00 ± 32. 91）,（ 79. 33 ± 20. 74） vs（ 208. 67 ± 73. 24）,all P〈0. 01]. After siRNA transfection,the Wip1 protein expression was down-regulated,and the cells proliferation was inhibited（ inhibition rate of cell proliferation was between 12. 37%- 40. 81%）. The cell migration distances at 0- 24 h,24- 48 h,48- 72 h after transfection in NC group were more than

关 键 词：肝癌 磷酸酶1基因 质粒转染 小干扰核糖核酸 细胞生物学特性 

分 类 号：R735.7[医药卫生—肿瘤]