Research Article - Biomedical Research (2017) Volume 28, Issue 21
NET-1 knockdown inhibits proliferation and promotes apoptosis of hepatocellular carcinoma cells by regulating apoptosis-related proteins and the PI3K/Akt signaling pathway
Background: Neuroepithelial transforming gene-1 (NET-1) is a guanine nucleotide exchange factor that activates Rho family proteins. We aimed to evaluate the effect of siRNA-mediated knockdown of NET-1 on proliferation and apoptosis of hepatocellular carcinoma (HCC) cells and the underlying mechanisms.
Material and Methods: NET-1 mRNA and protein levels were detected in four HCC cell lines MHCC97- L, MHCC97-H, SMCC7721, and HepG2, and the normal liver cell line L-02 using RT-PCR and western blot. Cell proliferation was evaluated by the CCK-8 assay, and apoptosis was assessed by flow cytometry. Protein levels of apoptosis-related proteins and PI3K/Akt pathway proteins were evaluated by western blot.
Results: NET-1 levels were significantly higher in the four HCC cell lines than those in the normal liver cell line L-02; the highest levels were observed in MHCC97-H cells. Knockdown of NET-1 by siRNA inhibited proliferation and promoted apoptosis of HCC cells. In addition, NET-1 knockdown decreased Bax and cyclin D1 expression, but increased Bcl-2 and caspase-3 levels in HCC cells. The PI3K/Akt pathway was blocked by knockdown of NET-1.
Conclusion: NET-1 knockdown inhibits proliferation and promotes apoptosis of HCC cells by regulating the expression of apoptosis-related proteins including Bax, Bcl-2, cyclin D1, and caspase-3, and the PI3K/Akt pathway.Author(s): Mingchun Wang, Xiangjun Sun, Yongjun Jiang, Maogang Chen