慢病毒介导靶向P27RF-Rho基因沉默对肝癌细胞侵袭性的影响  

Influence of lentiviral-mediated P27RF-Rho gene silence in invasion of liver cancer cells

作　　者：马强[1] 谢淑丽[1] 王广义[1] 邢光远 杨耀群 吕国悦[1] 

机构地区：[1]吉林大学第一医院肝胆胰外科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2016年第2期260-265,I0004,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(20130413021GH)

摘　　要：目的:探讨慢病毒靶向介导技术沉默P27RF-Rho基因,阐述其对肝癌细胞侵袭性的影响。方法:构建P27RF-Rho RNAi慢病毒。慢病毒感染肝癌细胞BEL7402。实验分为P27RF-Rho-siRNA实验组、Scramble-siRNA阴性对照组和BEL7402空白对照组。Western bloting法检测P27RF-Rho基因沉默效果及肝癌相关蛋白RhoA、RhoC、VEGF、P53和PTEN表达水平;明胶酶谱分析肿瘤侵袭相关基质金属蛋白酶(MMPs)活性;细胞划痕和体外Transwell小室侵袭实验比对细胞迁移和侵袭能力的改变。结果:Western blotting法检测,P27RF-Rho-siRNA实验组BEL7402中P27RF-Rho、RhoA、RhoC和VEGF蛋白表达水平明显低于2个对照组(P<0.05),P53和PTEN表达水平高于2个对照组(P<0.05)。明胶酶谱,P27RF-Rho-siRNA实验组MMPs活性明显降低(P<0.01)。划痕实验,P27RF-Rho-siRNA实验组细胞迁移距离明显低于2个对照组(P<0.01)。实验组穿过Transwell小室的平均细胞数明显少于2个对照组(P<0.01)。结论:沉默P27RF-Rho基因可以降低肝癌细胞BEL7402的侵袭和迁移能力。Objective： To investigate the silencing of P27RF-Rho gene with lenvirus targeting mediated technique, and to clarify its influence in the invasion of liver cancer cells. Methods： The P27RF-Rho RNAi lentivirus was constructed. The liver cancer BEL7402 cells were infected with lentivirus. The experiment was divided into P27RF Rho-siRNA group, scramble - siRNA group and BEL7402 group. The effect of silencing P27RF-Rho gene and the expression levels of hepatocellular carcinoma （HCC） associated proteins RhoA, RhoC, VEGF, P53 and PTEN were detected; the activities of matrix metalloproteinase （MMPs） associated with tumor invasion were analyzed by Gelatin zymography; the variation of transfer ability and invasion abilities were compared by Wound healing assay experiment and Transwell experiment. Results： The Western blotting results showed the expression levels of P27RF - Rho, RhoA, RhoC, and VEGF proteins in the BEL7402 cells in experiment group were significantly lower than those in two control groups （P〈0.05）, and the expression levels of P53 and PTEN were higher than those in two control groups （P〈0.05）. The results of Gelatin zymography showed the activities of MMPs in experiment group were significantly lower than those in two control groups （P〈 0.01）; Wound healing assay showed that the migration ability of the BEL7402 cells in experiment group was significantly inhibited （P〈0. 01） ; the number of cells passed through the Transwell Chambers in experiment group was significantly less than those in two control groups （P〈0.01）. Conclusion： Silenceing P27RF-Rho can weaken the invasion ability and migration ability of human HCC BELT402 cells.

关 键 词：P27RF-Rho 慢病毒 肝癌细胞 侵袭和迁移 

分 类 号：R735.7[医药卫生—肿瘤]