[Effect of hepatitis B virus X gene on the apoptosis of HepG2 cells].
OBJECTIVE
To investigate the effect of hepatitis B virus X gene on the apoptosis in X gene-transfected HepG2 cells.


METHODS
HBX gene eukaryon expression vector pcDNA3.1-X was transfected into HepG2 cells by lipid-mediated transfection to establish HepG2/HBX cell model for HBX expression. HepG2 transfected with pcDNA3.1 was used as controls. At 24 h, 48 h, 72 h and 96 h after transfection, cell apoptotic rates were detected by flow cytometry. RT-PCR and Western blot were applied to evaluate the expression levels of HBX, Fas, FasL, and the levels of p-JNK and p-c-Jun protein.


RESULTS
HBX mRNA was detected in HepG2/ HBX cells 24 h after transfection, and the expression of HBX mRNA level was gradually up-regulated after transfection (P < 0.05); whereas no expression of HBX gene in the control cells. At 24 h, 48 h, 72 h and 96 h after transfection, all of the apoptotic rate in HepG2/HBX group were much higher than those in the controls (P < 0.05). The expression levels of Fas and FasL protein as well as the levels of p-JNK and p-c-Jun protein were significantly higher than those in the controls (P < 0.05). A positive correlationship was observed between the expression of HBX mRNA level and the hepatocyte apoptotic rate (P < 0.05), the same correlation of HBX mRNA level with the levels of Fas, FasL, p-JNK and p-c-Jun were also observed. (P < 0.05).


CONCLUSIONS
HBX can induce hepatocyte apoptosis by up-regulating the levels of p-JNK and p-c-Jun to promote the expression of FasL.
Tang
Ren-Xian
RX
Department of Biological Pathogens, Xuzhou Medical College, Xuzhou 221002, China.
Shi
Hong-Li
HL
Fan
Bao-Feng
BF
You
Hong-Juan
HJ
Liu
Xiao-Mei
XM
Zheng
Kui-Yang
KY
D017209:Apoptosis / Q000235:genetics*; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D012333:RNA, Messenger; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein