CYP450蛋白在HepG2细胞与小鼠原代肝细胞中的表达差异

The Differential Protein Expression of CYP450 between HepG2 Cells and Mouse Primary Hepatocytes

目的:探讨CYP450蛋白在HepG2细胞与小鼠原代肝细胞中的表达差异。方法:采用改良的两步灌流法分离小鼠原代肝细胞后培养至36 h、同时培养HepG2细胞至对数生长期使用RIPA裂解液提取2种细胞的总蛋白;采用免疫印迹(Western blot)方法检测2种细胞中CYP2E1、CYP2C19、CYP2C9、CYP2D6、CYP3A4及CYP1A2的蛋白表达。结果:在HepG2细胞中未检测到CYP2E1、CYP2C19蛋白表达,但小鼠原代肝细胞中表达显著,2组比较差异有统计学意义(P<0.01);小鼠原代肝细胞中CYP2C9、CYP2D6、CYP3A4及CYP1A2的表达量分别约为HepG2细胞中的7.9倍、2.8倍、17.9倍及10.8倍,差异有统计学意义(P<0.01)。结论:小鼠原代肝细胞CYP450蛋白表达丰度较高,而HepG2细胞CYP450蛋白表达水平较低。

Objective:To investigate the difference of CYP450 protein expression between HepG2 cells and mouse primary hepatocytes.Methods:Mouse primary hepatocytes were isolated by a modified two-step perfusion method and cultured for 36 h,while HepG2 cells were cultured to logarithmic growth phase.RIPA lysate was used to extract the proteins from HepG2 cells and mouse primary hepatocytes.The protein expression levels of CYP2E1,CYP2C19,CYP2C9,CYP2D6,CYP3A4 and CYP1A2 was detected by Western blot.Results:CYP2E1 and CYP2C19 were detected in mouse primary hepatocytes expression,but not in HepG2 cells(P<0.01).The expression levels of CYP2C9,CYP2D6,CYP3A4,and CYP1A2 in mouse primary hepatocytes were approximately 7.9-fold,2.8-fold,17.9-fold,10.8-fold of HepG2 cells(P<0.01).Conclusion:CYP450 protein expression levels are high in mouse primary hepatocytes,but low in HepG2 cells.