沉默KIAA1429通过抑制Sirt1调控肝癌细胞的上皮间质转化

Silencing KIAA1429 regulates epithelial mesenchymal transformation of hepatoma cells by inhibiting Sirt1

目的研究沉默KIAA1429通过抑制Sirt1调控肝癌细胞的上皮间质转化(EMT)的作用。方法收集2007年9月至2014年1月首都医科大学宣武医院及首都医科大学附属北京佑安医院手术切除的肝癌组织及癌旁组织,培养肝癌细胞株SMMC-7721、HepG2、BEL-7402及人正常肝细胞株HL-7702,检测组织样本及细胞样本中KIAA1429、Sirt1的mRNA表达水平。将HepG2细胞分为NC-siRNA组、KIAA1429-siRNA组、KIAA1429-siRNA+Sirt1过表达组,转染siRNA或Sirt1表达质粒。检测细胞迁移率、KIAA1429、Sirt1及EMT标志基因E-cadherin、N-cadherin、Vimentin的mRNA表达水平。结果肝癌组织中KIAA1429、Sirt1的mRNA表达水平均高于癌旁组织,差异均有统计学意义(P<0.05),且KIAA1429的mRNA表达水平与Sirt1的mRNA表达水平呈正相关;肝癌细胞株SMMC-7721、HepG2、BEL-7402中KIAA1429、Sirt1的mRNA表达水平均高于人正常肝细胞株HL-7702,差异均有统计学意义(P<0.05);HepG2细胞中KIAA1429、Sirt1的mRNA表达水平均高于SMMC-7721、BEL-7402细胞,差异均有统计学意义(P<0.05)。KIAA1429-siRNA组HepG2细胞中KIAA1429、Sirt1、N-cadherin、Vimentin的mRNA表达水平及细胞迁移率均低于NC-siRNA组,E-cadherin的mRNA表达水平高于NC-siRNA组,差异均有统计学意义(P<0.05);KIAA1429-siRNA+Sirt1过表达组HepG2细胞中Sirt1、N-cadherin、Vimentin的mRNA表达水平及细胞迁移率均高于KIAA1429-siRNA组,E-cadherin的mRNA表达水平低于KIAA1429-siRNA组,差异均有统计学意义(P<0.05)。结论沉默KIAA1429通过抑制Sirt1表达的方式抑制肝癌细胞的EMT及迁移。

Objective To study the effect of silencing KIAA1429 regulates epithelial mesenchymal transformation(EMT)of hepatoma cells by inhibiting Sirt1.Methods Hepatocellular carcinoma tissues and adjacent tissues resected in Xuanwu Hospital and Beijing You'an Hospital,Capital Medical University from September 2007 to January 2014 were collected.Liver cancer cell lines SMMC-7721,HepG2,BEL-7402 and human normal liver cell line HL-7702 were cultured.The mRNA expression levels of KIAA1429 and Sirt1 in tissue and cell samples were detected.HepG2 cells were divided into NC-siRNA group,KIAA1429-siRNA group and KIAA1429 siRNA+Sirt1 overexpression group,then siRNA or Sirt1 expression plasmid was transfected.The cell mobility and the mRNA expression levels of KIAA1429,Sirt1 and EMT marker genes E-cadherin,N-cadherin and Vimentin were detected.Results The mRNA expression levels of KIAA1429 and Sirt1 in hepatocellular carcinoma were higher than those in adjacent tissues,the differences were statistically significant(P<0.05),and the mRNA expression level of KIAA1429 was positively correlated with that of Sirt1.The mRNA expression levels of KIAA1429 and Sirt1 in hepatoma cell lines SMMC-7721,HepG2 and BEL-7402 were higher than those in human normal hepatocyte line HL-7702,the differences were statistically significant(P<0.05);and the mRNA expression levels of KIAA1429 and Sirt1 in HepG2 cells were higher than those in SMMC-7721 and BEL-7402 cells,the differences were statistically significant(P<0.05).The cell mobility and the mRNA expression level of KIAA1429,Sirt1,N-cadherin and Vimentin in HepG2 cells of KIAA1429-siRNA group were lower than those in NC-siRNA group,and the mRNA expression level of E-cadherin was higher than that in NC-siRNA group,the differences were statistically significant(P<0.05);the cell mobility and the mRNA expression level of Sirt1,N-cadherin and Vimentin in HepG2 cells of KIAA1429-siRNA+Sirt1 overexpression group were higher than those in KIAA1429-siRNA group,and the mRNA expression level of E-cadherin was lower than that in KIAA1429-siRNA group,the differences were statistically significant(P<0.05).Conclusion Silencing KIAA1429 inhibits EMT and migration of hepatoma cells by inhibiting Sirt1 expression.