HBx及其突变体X17-3对Hippo信号途径表达的影响

HBx and its mutant X17-3 regulates YAP through Hippo signaling pathway in human hepatic L02 cells

目的初步探寻HBx及其不同突变体表达与Hippo信号途径的关系,以及对细胞凋亡的影响。方法 HBx及其突变体载体转染人正常肝细胞系L02,转染48 h后,提取总蛋白,Western blot检测细胞中Hippo信号途径MST1、YAP(yes-associated protein)的表达,磷酸化和去磷酸化的情况。采用Annexin V/PI标记流式细胞术检测细胞凋亡。结果HBx及其突变体载体转染L02细胞48 h后,Western blot结果显示细胞MST1表达下调,p-MST1/2表达上升(P<0.05);YAP表达上升,p-YAP表达下调(P<0.05)。Annexin V/PI标记法流式细胞术结果显示HBx及其突变体能促进L02细胞发生凋亡(P<0.05),其介导的细胞早期凋亡尤为明显。结论 HBx通过Hippo信号通路途径调控下游致癌基因YAP的表达。结合HBx介导的L02细胞凋亡这一结果说明HBx可能通过多种途径调节细胞周期。

ling pathway, and Object ve To investigate the the effects of HBx on cell cycle relationship of HBx and its and apoptosis. Methods mutant X17-3 with Hippo signaL02 cells were transfected with HBx and its mutant, respectively. After 48 h of incubation, the expression levels as well as the phosphorylation status of MST1/2 and yes-associated protein （YAP） were detected by Western blotting. The apoptosis of L02 cells was examined with flow cytometry by Annexin V/PI staining. Results After 48 hours＇ transfection, Western blotting showed that the expression levels of p-MST1/2 and YAP were up-regulated, and those of MST1/2 and p-YAP were down-regulated （P 〈 0. 05 ）. The results of flow cytometry indicated that the transfection of HBx and X17-3 resulted in more apoptotic cells （ P 〈 0. 05 ）, especially in early stage of apoptosis. Conclusion HBx regulates its downstream oncogene, YAP through Hippo signaling pathway. Its also regulates the cell cycle via several ways.