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Apoptosis and its pathway in X gene-transfected HepG_2 cells

World Journal of Gastroenterology 2005年第28期11卷 4326-4331页

作者： Na Lin Hong-Ying chen Dan Li Sheng-Jun Zhang Zhi-Xin cheng Xiao-Zhong Wang Department of Gastroenterology Affiliated Union Hospital Fujian Medical University Fuzhou 350001 Fujian Province China

AIM： To investigate the effect of hepatitis B virus （HBV） X gene on apoptosis and expressions of apoptosis factors in X gene-transfected HepG2 cells.METHODS： The HBV X gene eukaryon expression vector pcDNA3-X was transiently transfected into HepG2 cells by lipid-media transfection. Untransfected HepG2 and HepG2 transfected with pcDNA3 were used as controls. Expression of HBx in HepG2 was identified by RT-PcR. MTT and TUNEL were employed to measure proliferation and apoptosis of cells *** groups. Semi-quantified RT-PcR was used to evaluate the expression levels of Fas/FasL, Bax/Bcl-xL, and c-myc in each group.RESULTS： HBV X gene was transfected into HepG2 cells successfully. RT-PcR showed that HBx was only expressed in HepG2/pcDNA3-X cells, but not expressed in HepG2 and HepG2/pcDNA3 cells. Analyzed by MTT, cell proliferationcapacity was obviously lower in HepG2/pcDNA3-X cells （0.08910±0.003164） than in HepG2 （0.14410±0.004927） and HepG2/pcDNA3 cells （0.12150±0.007159）（P〈0.05 and P〈0.01）. Analyzed by TUNEL, cell apoptosis was much more in HepG2/pcDNA3-X cells （980/2 000） than HepG2 （420/2 000）, HepG2/pcDNA3 cells （520/2 000）（P〈0.05 and P〈0.01）. Evaluated by semi-quantified RT-PcR, the expression level of Fas/FasL was significantly higher in HepG2 cells transfected with HBx than in HepG2 and HepG2/pcDNA3 cells （P〈0.05 and P〈0.01）. Bax/Bcl-xL expression level was also elevated in HepG2/pcDNA3-X cells （P〈0.05 and P〈0.01）. Expression of c-myc was markedly higher in HepG2/pcDNA3-X cells than in HepG2 and HepG2/pcDNA3 cells （P〈0.05 and P〈0.01）.cONcLUSION： HBV X gene can impair cell proliferation capacity, improve cell apoptosis, and upregulate expression of apoptosis factors. The intervention of HBV X gene on the expression of apoptosis factors may be a possible mechanism responsible for the change in cell apoptosis and proliferation.

关键词： HBx Transfect HepG2 Apoptosis Fas FasL Bax Bcl-xL c-mvc

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