BAI Guangwen, SHEN Lihong, LU Yongliang, et al. Effect of hepatitis B virus X protein mediated by adenovirus on the differentiation of hepatic progenitor cells[J]. Journal of Chongqing Medical University, 2012,37(10):846-849.
BAI Guangwen, SHEN Lihong, LU Yongliang, et al. Effect of hepatitis B virus X protein mediated by adenovirus on the differentiation of hepatic progenitor cells[J]. Journal of Chongqing Medical University, 2012,37(10):846-849.DOI:
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.
关键词
乙肝病毒X蛋白肝前体细胞分化肿瘤干细胞
Keywords
hepatitis B virus X proteinhepatic progenitor celldifferentiationtumor stem cell