OCT4介导肝细胞去分化过程中巢蛋白的表达及意义探讨

Expression of nestin in OCT4-mediated dedifferentiation of hepatocytes and its significance

目的探讨OCT4介导的原代肝细胞去分化过程中巢蛋白(nestin)的表达及其意义。方法体外分离培养小鼠原代肝细胞。包装、浓缩表达OCT4的慢病毒并感染肝细胞。观察细胞的形态变化,用RT-PCR法监测nestin、白蛋白(albumin,ALB)、甲胎蛋白(alpha-fetoprotein,AFP)以及叉头框A2(FOXA2)的表达并进行定量分析。结果 OCT4慢病毒感染后,肝细胞内颗粒逐渐释出,并在7 d左右开始出现肝细胞增殖。在肝细胞ALB减弱,AFP、FOXA2增强的过程中,nestin从第3天持续至第9天,表达逐渐增强,从12 d起开始减弱,至18 d时上述基因的表达几近消失。结论 Nestin的表达伴随肝细胞去分化和增殖的过程。

Objective To study the expression of nestin in OCT4-mediated dedifferentiation of hepatocytes and its significance.Methods Mouse primary hepatocytes were isolated and cultured in vitro.Hepatocytes were infected with packaged and concentrated OCT4-lentivirus.Morphology of hepatocytes was observed.Expressions of nestin,ALB,AFP,and FOXA2 were detected by RT-PCR and quantitatively analyzed.Results After the hepatocytes were infected with OCT4-lentivirus,the intracellular particles were gradually released from hepatocytes and the hepatocytes began to proliferate from day 7,the nestin increased its expression from day 3 to day 7 and began to decrease from day 12 when the ALB reduced it expression while the AFP and FOXA2 increased their expressions.The expressions of these genes were hardly detected on day18 after the hepatocytes were infected with OCT4-lentivirus.Conclusion Nestin is expressed with the dedifferentiation and proliferation of hepatocytes.