跨膜丝氨酸蛋白酶4对肝细胞癌增殖侵袭作用的影响

摘要

目的 探讨跨膜丝氨酸蛋白酶4 (transmembrane protease serine 4,TMPRSS4)对肝细胞癌(hepatocellular carcinoma,HCC)增殖侵袭的作用.方法 构建并鉴定慢病毒转染TMPRSS4过表达的人肝细胞癌细胞系BEL-7402.Western blot及RT-PCR检测TMPRSS4的表达.MTT实验检测TMPRSS4对BEL-7402细胞增殖能力的影响.Transwell实验检测TMPRSS4对细胞侵袭性的影响.结果 RT-PCR及Western blot结果显示TMPRSS4过表达组细胞TMPRSS4蛋白及mRNA表达均显著高于对照组及空白载体转染组.MTT实验结果显示,慢病毒转染后,对照组、空白载体转染组和TMPRSS4过表达组细胞增殖能力无显著差异(P＞0.05).侵袭实验显示TMPRSS4过表达组细胞穿过Matrigel胶到达Transwell小室膜背面的细胞数为49.3±7.4,显著高于对照组(23.3±5.0)和空白载体转染组(19.3±3.2)(P＜0.05).结论 HCC细胞表达TMPRSS4.TMPRSS4过表达可促进HCC细胞的侵袭能力,但是不影响HCC细胞增殖.%Objective This study investigates the effects of transmembrane protease serine 4 (TMPRSS4) on the proliferation and invasion of hepatocellular carcinoma (HCC).Methods Stable TMPRSS4 over-expression cell lines were established using human HCC cells BEL-7402 which were transfected by lentiviral vectors.Expression of TMPRSS4 was measured by Western blot and RT PCR.Cell proliferation was measured by MTT analysis and HCC cell invasion was evaluated by tran swell invasion assay.Results RT-PCR and Western blot analysis showed that compared to control HCC cells (control group) and cells transfected with blank plasmid (blank plasmid group),the expression of TMPRSS4 in cells transfected with TMPRSS4 plasmid (TMPRSS4 plasmid group) was significantly increased.There were no significant differences among these three groups regarding the proliferation of HCC cells (P＞0.05).The number of cells in the TMPRSS4 plasmid group that passed through the transwell inserts was 49.3 ± 7.4,significantly higher than that of control group and blank plasmid group,which was 23.3 ± 5.0 and 19.3 ± 3.2 respectively (P＜0.05).Conclusion Therefore,the expression of TMPRSS4 can be detected in HCC,and over-expression of TMPRSS4 promotes the invasion of HCC but has no effect on cell proliferation.