乙肝病毒X蛋白对肝前体细胞分化的影响

Effect of hepatitis B virus X protein mediated by adenovirus on the differentiation of hepatic progenitor cells

摘要:

目的：检测乙肝病毒X（Hepatitis B virus X，HBx）蛋白对肝前体细胞分化的影响，以探讨HBx蛋白能否促进肝干细胞的恶性转化。方法：以腺病毒Ad-HBx和Ａd-GFP感染肝前体细胞株Hp14-19，RT-PCR和Western blot证实HBx蛋白在肝前体细胞内的表达。RT-qPCR和免疫荧光检测早期和晚期分化指标的变化，并于诱导后11 d用PAS染色观察细胞的分化成熟程度。结果：Ad-HBx能有效感染肝前体细胞株Hp14-19并表达。与对照组（感染Ad-GFP）相比，实验组（感染Ad-HBx）细胞分化早期指标穿膜蛋白（Delta like homolog，DLK）、肌酸激酶19（Creatine kinase19，CK19）和甲胎蛋白（Alpha fetal protein，AFP）表达降低减慢，而晚期指标CK18和白蛋白（Albumin，ALB）表达升高不明显。PAS染色阳性率降低。差异具有统计学意义（P<0.05）。结论：HBx可抑制肝前体细胞的分化，这可能是最终导致肝前体细胞恶性转化的原因之一。

Abstract:

Objective：To detect the effect of hepatitis B virus X（HBx） protein on the differentiation of hepatic progenitor cell and to explore whether HBx protein can promote the malignant transformation of hepatic stem cell. Methods：The hepatic progenitor cell Hp14-19 was infected with adenovirus Ad-HBx and Ad-GFP. The expressions of HBx protein in hepatic progenitor cell were con－firmed by RT-PCR and Western blot. The changes of differentiation marker alpha fetal protein（AFP），delta like homolog（DLK），creatine kinase19（CK19），CK18 and albumin（ALB） in the hepatic progenitor cell treated with HBx were evaluated by RT-qPCR and immunofluorescence. The morphological change was observed by PAS staining at 11 d after the inducement. Results：HBx was ex－pressed in the hepatic progenitor cell Hp14-19 infected with Ad-HBx. Compared with those in control group（infected with Ad-GFP），the expressions of early marker DLK，CK19 and AFP were decreased slowly while later marker CK18，ALB were increased slowly in experiment group（infected with Ad-HBx）. PAS staining results showed that HBx inhibited glycogen synthesis in HP14.5 cells. Con－clusions：HBx can inhibit the differentiation of hepatic progenitor cell，which maybe involved in the tumorigenesis of HBx-mediated liver cancer.