摘要
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-β_1、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。
Objective：To investigate the role and molecular mechanism of Na~＋/Ca^（2＋） 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（MHCC97 H,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-β_1,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 EMTrelated proteins in HCC cells,and it is a potential target for the diagnosis and treatment of liver cancer.
出处
《中国临床医学》
2017年第6期873-878,共6页
Chinese Journal of Clinical Medicine
基金
国家自然科学基金面上项目(81572395)~~
关键词
钠钙交换器1
肝细胞肝癌
侵袭
增殖
上皮间质转化
Na^＋/Ca^2＋exchanger 1
hepatocellular carcinoma
invasion
proliferation
epithelial-mesenchymal transition
