钠钙交换器1对肝细胞肝癌增殖与侵袭的影响

摘要

Objective:To investigate the role and molecular mechanism of Na+/Ca2+ exchanger 1 (NCX1) in the invasion and proliferation of hepatocellular carcinoma (HCC),and to provide new therapeutic targets for the clinical diagnosis and treatment of hepatocellular carcinoma.Methods:Quantitative real-time PCR (qRT-PCR) and Western blotting were used to detect NCX1 mRNA and protein expression in different HCC cell lines.Lentiviral vectors pGC-SIL-GFP-shRNA-NCX1 and pGC-FU-GFP-NCX1 were constructed and transfected into hepatoma cells.Cell migration and invasion experiments were used to observe the effect of NCX1 on the migration and invasion of HCC cells;cell proliferation assay was used to observe the effect of NCX1 on the proliferation of hepatoma cells;Western blotting and ELISA were used to detect the expressions of NCX1-related proteins.Results:The expression of NCX1 in high metastatic potential HCC cell lines (MHCC97H,HCCLM3) was higher than that of low metastatic potential HCC cell lines (HepG2,Huh7,SMMC-7721),and the difference was statistically significant (P＜0.05).After transfection of NCX1,the invasion and proliferation ability of HCC cells increased significantly (P＜0.05);the secretion of cytokines (TGF-β1,IL-6 and TNF-α) increased significantly (P＜0.05);the expressions of epithelial-mesenchymal transition (EMT) related proteins in HCC cells were significantly increased (P＜0.05).Conclusions:NCX1 can promote the growth,invasion and metastasis of hepatocellular carcinoma by increasing the expressions of EMT-related proteins in HCC cells,and it is a potential target for the diagnosis and treatment of liver cancer.%目的:探讨钠钙交换器1 (NCX1)在肝细胞肝癌(简称肝癌)侵袭和增殖中的作用及分子机制,为肝癌的临床诊治提供新思路.方法:采用荧光定量PCR(qRT-PCR)、Western印迹法检测不同肝癌细胞系中NCX1的表达情况.构建NCX1干扰慢病毒载体和过表达慢病毒载体,并转染肝癌细胞.采用细胞迁移、侵袭实验观察NCX1对肝癌细胞迁移、侵袭的影响;采用细胞增殖实验观察NCX1对肝癌细胞增殖的影响;ELISA、Western印迹法检测NCX1相关蛋白的表达.结果:高转移潜能肝癌细胞系(MHCC97H、HCCLM3) NCX1表达高于低转移潜能肝癌细胞系(HepG2、Huh7、SMMC-7721),差异有统计学意义(P＜0.05).转染NCX1后,肝癌细胞侵袭及增殖能力明显增加(P＜0.05);细胞因子(TGF-β1、IL-6、TNF-α)分泌明显增多(P＜0.05);肝癌上皮间质转化(EMT)相关蛋白表达明显升高(P＜0.05).结论:NCX1体外可能通过升高肝癌细胞EMT相关蛋白表达促进肝癌生长、侵袭及转移,是潜在的肝癌诊治靶标.