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Exogenous phosphatidylethanolamine induces apoptosis of human hepatoma hepg2 cells via the bcl-2/bax pathway

World Journal of Gastroenterology 2009年第14期15卷 1751-1758页

作者： Yu Yao Chen Huang Zong-Fang Li Ai-Ying Wang Li-Ying Liu Xiao-Ge Zhao Yu Luo Lei Ni Wang-Gang Zhang Tu-Sheng Song Department of Oncology First Affiliated Hospital of Medical College of Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China Department of Genetics and Molecular Biology Medical School Xi'an Jiaotong University/Key Laboratory of Environment and Genes Related to Diseases Ministry of Education Xi'an 710061 Shaanxi Province China Second Affiliated Hospital of Xi'an Jiaotong University/Key Laboratory of Environment and Genes Related to Diseases Ministry of Education Xi'an 710004 Shaanxi Province China

AIM: To investigate the signaling pathways implicated in phosphatidylethanolamine (PE)-induced apoptosis of human hepatoma hepg2 cells. METHODS: Inhibitory effects of PE on human hepatoma hepg2 cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. cell cycle, apoptosis and mitochondrial transmembrane potential (ΔΨm) were analyzed by flow cytometry. Immunocytochemical assay and Western blotting were used to examine Bcl-2, Bax and caspase-3 protein levels in hepg2 cells treated with PE. RESULTS: PE inhibited the growth of hepg2 cells in a doseand timedependent manner. It did notaffect the cell cycle, but induced apoptosis. PE significantly decreased ΔΨm at 0.25, 0.5 and 1 mmol/L, respectively, suggesting that PE induces cell apoptosis by decreasing the mitochondrial transmembrane potential. The Bcl-2 expression level induced by different concentrations of PE was lower than that in control groups. However, the Bax expression level induced by PE was higher than that in the control group. Meanwhile, PE increased the caspase-3 expression in a doseand time-dependent manner. CONCLUSION: Exogenous PE induces apoptosis of human hepatoma hepg2 cells via the bcl-2/bax pathway.

关键词： Apoptosis Bcl-2 Bax Caspase-3 Phosphatidylethanolamine human hepatoma hepg2 cell

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Molecular mechanisms of apoptosis induced by Scorpio water extract in human hepatoma hepg2 cells

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World Journal of Gastroenterology 2005年第7期11卷 943-947页

作者： Kang-Beom Kwon Eun-Kyung Kim Jung-Gook Lim Eun-Sil Jeong Byung-Cheul Shin Young-Se Jeon Kang-San Kim Eun-A Seo Do-Gon Ryu Department of Physiology Department of Rehabilitation Medicine Department of Internal Medicine Department of Food and Nutrition

AIM: To clarify the mechanism underlying the anti-mutagenic and anti-cancer activities of Scorpio water extract (SWE). METHODS: human hepatoma hepg2 cells were incubated with various concentrations of SWE. After 24-h incubation, cytotoxicity and apoptosis evaluations were determined by MTT and DNA fragmentation assay, respectively. After treatment with SWE, mitochondrial membrane potential (MMP) was determined by measuring the retention of the dye 3,3'-dihexyloxacarbocyanine (DiOC6(3)) and the protein expression including cytochrome C and poly-(ADPribose) polymerase (PARP) were measured by Western blotting. Caspase-3 and -9 enzyme activities were measured using specific fluorescence dyes such as Ac-DEVD-AFC and Ac-LEHD-AFC. RESULTS: We found that treatment with SWE induced apoptosis as confirmed by discontinuous DNA fragmentation in cultured human hepatoma hepg2 cells. Our investigation also showed that SWE-induced apoptosis of hepg2 cells were associated with intracellular events including disruption of MMP, increased translocation of cytochrome C from mitochondria to cytosol, activation of caspase-3, and PARP. Pre-treatment of N-acetyl-Asp-Glu-Val-Asp-CHO (Ac-DEVD-CHO), a caspase-3 specific inhibitor, or cyclosporin A (CsA), an inhibitor of MMP disruption, completely abolished SWE-induced DNA fragmentation. CONCLUSION: These results suggest that SWE possibly causes mitochondrial damage, leading to cytochrome C release into cytosol and activation of caspases resulting in PARP cleavage and execution of apoptotic cell death in hepg2 cells. These results further suggest that Scorpio may be a valuable agent of therapeutic intervention of human hepatomas.

关键词： Scorpio human hepatoma hepg2 cell Apoptosis

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