Department of Pharmacy, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China.
Biomed Res Int. 2013;2013:434561. doi: 10.1155/2013/434561. Epub 2013 Jul 14.
This study aims to investigate the effects of ADAM10 expression on metastasis and invasiveness of human hepatocellular carcinoma HepG2 cells.
The HepG2 cells were transfected with medium only, the empty vector, the control siRNA, or siRNA against ADAM10, respectively. Cell migration assay and Transwell invasiveness assay were performed to detect the effects of ADAM10 knockdown on migration and invasiveness of HepG2 cells. Western blotting and real-time RT PCR were performed to investigate the effects of knock-down of ADAM10 on protein and mRNA levels of E-cadherin gene.
Cell migration and invasiveness of HepG2 cells transfected with ADAM10 siRNA were significantly decreased, when compared with the cells transfected with the control siRNA, suggesting that the downregulation of ADAM10 expression inhibits cell migration and invasiveness. The Western blotting results suggest that the down-regulation of ADAM10 expression increases E-cadherin protein levels. The real-time RT-PCR results indicated that the mRNA level of E-cadherin is not detectably affected by the knock-down of ADAM10 gene.
Expression of ADAM10 may be related to cell migration and invasiveness of human hepatocellular carcinoma HepG2 cells via a mechanism related to E-cadherin.
本研究旨在探讨 ADAM10 表达对人肝癌 HepG2 细胞转移和侵袭的影响。
分别用普通培养基、空载体、对照 siRNA 或针对 ADAM10 的 siRNA 转染 HepG2 细胞。通过细胞迁移实验和 Transwell 侵袭实验检测 ADAM10 敲低对 HepG2 细胞迁移和侵袭的影响。通过 Western blot 和实时 RT-PCR 检测 ADAM10 敲低对 E-钙黏蛋白基因蛋白和 mRNA 水平的影响。
与转染对照 siRNA 的细胞相比,转染 ADAM10 siRNA 的 HepG2 细胞的迁移和侵袭能力显著降低,提示 ADAM10 表达下调抑制细胞迁移和侵袭。Western blot 结果提示 ADAM10 表达下调增加 E-钙黏蛋白蛋白水平。实时 RT-PCR 结果表明 ADAM10 基因敲低对 E-钙黏蛋白的 mRNA 水平没有明显影响。
ADAM10 的表达可能通过与 E-钙黏蛋白相关的机制与人类肝癌 HepG2 细胞的迁移和侵袭有关。