The anti-apoptotic effect of cytoplasmic alpha-fetoprotein in hepatoma cells induced by all-trans retinoic acid involves activation of the PI3K/AKT signaling pathway.
- VernacularTitle:甲胎蛋白激活PI3K/AKT信号促使肝癌Bel 7402细胞抗全反式维甲酸诱导凋亡
- Author:
Ming-Yue ZHU
1
;
Jun-Li GUO
;
Hua XIA
;
Wei LI
;
Yan LU
;
Xu DONG
;
Yi CHEN
;
Shi-Gan FU
;
Xie-Ju XIE
;
Feng-Sen LI
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Blotting, Western; Carcinoma, Hepatocellular; metabolism; Cell Line, Tumor; Cytoplasm; Humans; Immunoprecipitation; Liver Neoplasms; metabolism; PTEN Phosphohydrolase; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; RNA Interference; RNA, Small Interfering; Signal Transduction; drug effects; Transfection; Tretinoin; pharmacology; alpha-Fetoproteins; metabolism
- From: Chinese Journal of Hepatology 2014;22(11):837-842
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of alpha-fetoprotein (AFP) on transduction of the PI3K/ AKT signal in hepatocellular carcinoma cells and the role played by AFP in resistance to cytotoxicity of all-trans retinoic acid (ATRA).
METHODSThe effects of ATRA of human liver cancer cells was assessed using the BEL-7402 cell line with the MTT assay (to evaluate proliferation), microscopy (to evaluate morphology), flow cytometry (to evaluate apoptosis), laser confocal microscopy and coimmunoprecipitation (co-IP; to evaluate co-localization and interaction of AFP with PTEN), Western blotting (to evaluate expression of phosphorylated-protein kinase B (pAKT) and Src, and RNA interference (RNAi)-mediated knockdown of AFP. Finally, application of the PI3K-specific inhibitor Ly294002 was used to monitor the influence of AFP in transduction of the PI3K signal pathway.
RESULTSThe human hepatoma cell line BEL-7402 were resistant to ATRA cytotoxicity. PTEN and AFP co-localized in the cytoplasm, and co-IP indicated that AFP interacts with PTEN in BEL-7402 cells.RNAi knockdown of AFP expression led to reduced growth of BEL-7402 cells.BEL-7402 cells transfected with AFP-short interfering (si)RNA vectors showed enhanced sensitivity to ATRA and reduced expression of pAKT(Ser473) and Src; Ly294002 reduced the role of AFP in stimulating expression of pAKT(Ser473) and Src.
CONCLUSIONAFP can activate transduction of the PI3K/AKT signal, and expression of AFP in hepatoma cells is a pivotal event for resisting ATRA-induced apoptosis.
