HBV转染及缺氧应激对HepG2细胞VEGF表达的影响

EFFECTS OF HBV INFECTION AND HYPOXIA ON THE EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN HEPATOMA CELLS

目的探讨HBV转染及缺氧应激对HepG2细胞VEGF表达的影响。方法分别在常氧与缺氧条件下体外培养肝癌细胞系HepG2细胞及HepG2.2.15细胞,用倒置显微镜观察两株细胞形态学变化,在不同时间点(2、6、12、24、48h)检测二者在不同条件下VEGFmRNA及蛋白表达的差异。用Westen-Blot法检测VEGF蛋白表达、半定量RT-PCR技术检测细胞中VEGFmRNA相对表达量。结果各组VEGFmRNA及其蛋白表达均为阳性;HepG2.2.15细胞VEGFmRNA及其蛋白表达高于HepG2细胞(P<0.05);缺氧各组VEGFmRNA及其蛋白相对表达量随时间的增加与常氧组VEGFmRNA及其蛋白相对表达量呈正相关(两种方法测得r值分别为0.879与0.821,0.823与0.822)。结论 HBV感染及缺氧均能够刺激HepG2细胞表达VEGF;并且在缺氧刺激下,HepG2细胞VEGF表达增加,表达量与时间呈正相关。

Objective To explore the effects of HBV infection and hypoxia on the expression of Vascular Endothelial Growth Factor（VEGF） in Hepatoma cells.Methods HepG2 cells and HepG2.2.15 cells were cultured in vitro in normoxic and hypoxic conditions. The cell morphology was observed through the inverted light micro-scope.The change of VEGF mRNA and its protein was detected under different conditions within different time points（2h,6h,12h,24h,48h）.The relative expression levels of VEGF mRNA and protein intwo cells were detected by reverse transcription polymerase chain reaction（RT-PCR） and Western-Blot respectively. Results Expression of VEGF mRNA and its protein in every group are positive.The expression level of VEGF mRNA and its protein in HepG 2.2.15 cell is significantly higher than in HepG2 cell（P〈0.05）.Increased with time,the expression of VEGF mRNA and its protein in each of hypoxic group was positively correlated with which in normoxic group（r values were 0.879,0.821 and 0.823,0.822,respectively）.Conclusion HBV infection and hypoxia can stimulate the expression of VEGF in HepG2 cells.The expression of VEGF in HepG2 cells is increased with the time under hypoxic exposure.