摘要
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.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第16期1704-1707,共4页
Journal of Third Military Medical University
基金
国家重大传染病专项课题(2012ZX10002006)~~