TITLE:
TRAF6 Affects RAC1 Expression and Apoptosis in SK-Hep1 Cells
AUTHORS:
Shanlei Zhang, Changshi Qian, Xiaochen Liu, Shengjun Piao, Xinglin Jin
KEYWORDS:
Apoptosis, RAC1, TRAF6, FADD, RIP1, SK-Hep1 Cells
JOURNAL NAME:
Chinese Medicine,
Vol.9 No.4,
December
27,
2018
ABSTRACT: RAC1 is a small-molecule G
protein that regulates multiple cell cycle, cytoskeletal reorganization, cell
migration, and apoptosis. FADD-dependent TRAIL can promote tumor metastasis
through RAC1 and PI3K, and down-regulating RAC1 expression can reduce
FasL-induced apoptosis. In addition, RIP1 bound to GTP acts as an activating
protein for RAC1 and is involved in cytoskeletal reorganization. TRAF6 promotes
migration and metastasis by regulating the RAS pathway in tumors. Thus, it is
necessary to understand the interaction between RAC1 and TRAF6 as well as FADD and RIP1. In this study, we cultured hepatoma SK-Hep1 cells in
vitro, specifically blocked the necroptosis pathway with Nec-1, and
silenced FADD, RIP1 and TRAF6 gene expression using RNAi technology. At the
same time, the expression of RAC1 was evaluated separately using RT-PCR and
Western blot. The hepatoma SK-Hep1 cells survival rate was highest when the
concentration of Nec-1 was 60 μM and the concentration of Z-vad-fmk was 20 μM.
And the apoptosis rate of the transfected RAC1 siRNA cells was 3.59% compared
with transfected siRNA cells 10.01% which was significantly decreased (P