HBV—X蛋白对人肝癌细胞株SMMC-7721恶性表型的影响  

Effects of HBV-X on the malignant phenotypes of human hepatocellular carcinoma cell line SMMC- 7721

作　　者：孙长宇[1] 吴阳[2] 薛建峰[2] 杨观瑞 

机构地区：[1]郑州大学第一附属医院感染科,450052 [2]郑州大学第一附属医院普外科,450052 [3]河南省医药科学研究院

出　　处：《中华实验外科杂志》2010年第7期892-894,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察HBV-X蛋白对低侵袭性人肝癌细胞株SMMC-7721恶性表型的影响.方法 pcDNA3.1（-）-X重组质粒转染SMMC-7721细胞,以空质粒转染作对照,用逆转录-聚合酶链反应（RT-PCR）,Western blot检测X蛋白表达水平,用酶联免疫吸附试验（ELISA）检测细胞培养上清液X蛋白、MMP-2、MMP-9、尿激酶纤溶酶原活化因子（uPA）水平,并用体外功能实验观察转染前后的恶性表型的变化.结果 重组质粒转染SMMC-7721后X蛋白表达明显升高,细胞培养上清液HBV-X蛋白为（13.02±2.12）μg/L,对照组为（4.07±0.37）mg/L MMP-2重组质粒转染组为（49.04±4.07）μg/L,对照组为（25.15±5.43）μg/L,MMP-9水平分别为（27.82±2.25）μg/L和（7.89±1.27）μg/L uPA重组质粒转染组水平明显高于空质粒转染组,分别为（10.78±3.23）μg/L和（1.24±0.34）μg/L,t值分别为19.93,6.27 6.83,7.04,P值均〈0.01.体外功能实验提示SMMC-7721转染HBV-X重组质粒后细胞黏附、运动和侵袭能力明显增强,细胞黏附率为（85.33±14.78）%,对照组为（57.34±2.52）%,t=2.96,P〈0.05.运动试验透膜组胞数分别为（24.3±1.9）个和（12.3±2.5）个,t=4.89,P〈0.05.侵袭试验透膜细胞数分别为（18.2±3.2）个和（9.3±2.3）个,t=4.29,P〈0.05.而细胞增殖能力也明显增加.结论 HBV-X蛋白可能通过MMP-2、MMP-9、uPA分泌促进SMMC-7721的恶性表型.Objective To study the effects of HBV-X on the malignant phenotypes of human carcinoma cell line SMMC-7721. Methods Human hepatocellular carcinoma SMMC-7721 cells were transfect-ed with plasmid pcDNA3. 1（-）-X and cells transfected with a mock plasmid served as controls. HBV-X expression was detected by using reverse transcription-polymerase chain reaction （ RT-PCR） and Western blotting, and concentrations of HBV-X, MMP-2, MMP-9 and urinase plasminogen activator （uPA） were measured by enzyme-linked immunosorbent assay （ELISA）. A series of functional assays in vitro were used to monitor the changes of SMMC-7721 malignant phenotypes. Results HBV-X expression of SMMC-7721 cells was elevated after transfection. Concentrations of HBV-X, MMP-2, MMP-9 and uPA in the medium of SMMC-7721 cells after transfection were higher than those of the controls [（13.02 ± 2. 12）μg/L vs （4. 07 ±0.37）μg/L, （49.04±4.07）μg/L vs （25.15 ±5.43）μg/L, （27. 82 ±2. 25）μg/L vs （7.89± 1.27）μg/L, （10.78±3.23）μg/L vs （1.24±0.34）μg/L respectively,P〈0.01]. Functional assays ire vitro indicated that SMMC-7721 cells after transfection showed higher adhensive, migrant and invasive capabilities than those of the controls [cell adhesion rate： （85.33±14.78） % vs （57.34±2.52）% number of outer surface cells in migrant assay： 24.3±1.9 vs 12.3±2.5 cell number in the invasive assay： 18.2± 3.2 vs 9.3±2.3 respectively.P〈0.05]. Conclusion HBV-X might enhance the expression of MMP-2, MMP-9 and uPA to promote malignant phenotypes of SMMC-7721 cells.

关 键 词：癌 肝细胞 HBV-X蛋白 

分 类 号：R735.7[医药卫生—肿瘤]