摘要
目的探讨乙型肝炎病毒(HBV)对抑制素(PHB)表达的影响及PHB在肝细胞癌(HCC)细胞增殖和存活中的作用。方法应用实时荧光定量PCR技术和蛋白质印迹法检测13对HBV感染肝脏和正常肝脏及HepG2.2.15和HepG2细胞的PHB表达情况。收集7例慢性乙型肝炎患者抗病毒(替诺福韦酯)治疗前后的肝组织,采用RT-PCR技术和蛋白质印迹法检测PHB的表达。收集转染了Pcmv6-AC-GFP-PHB和对照载体的HepG2.2.15细胞。通过流式细胞仪分析DNA的含量。应用细胞增殖测定法检测每个细胞组的增殖水平。将转染了Pcmv6-AC-GFP-PHB的HepG2.2.15细胞和对照载体在无血清培养基中培养6 d。采用基于荧光激活细胞分选(FACS)的Annexin-V/碘化丙啶双重染色在指定的时间点检测细胞凋亡。组间比较采用Student t检验。结果与正常肝组织相比,HBV感染肝组织PHB表达下调(P<0.01);与HepG2细胞相比,HepG2.2.15细胞PHB表达显著下降(P<0.01)。抗病毒(替诺福韦酯)治疗后的肝组织PHB表达水平较治疗前明显上调(P<0.01)。与对照载体相比,转染了Pcmv6-AC-GFP-PHB的HepG2.2.15细胞增殖速率明显低于对照载体,转染了Pcmv6-AC-GFP-PHB载体的HepG2.2.15细胞的凋亡率显著高于对照(P<0.01)。结论HBV下调PHB表达从而促进HCC细胞的增殖和存活。
Objective To investigate the effect and role of the hepatitis B virus(HBV)on the expression of inhibin(PHB)in the proliferation and survival of hepatocellular carcinoma(HCC)cells.Methods The expression of PHB in 13 pairs of HBV-infected livers,normal livers and HepG2.2.15 and HepG2 cells was detected by real-time fluorescent quantitative PCR and Western blot.Liver tissues were collected from seven patients with chronic hepatitis B before and after antiviral(tenofovir)treatment,and the expression of PHB was detected by RT-PCR and Western blot.HepG2.2.15 cells were transfected with Pcmv6-AC-GFP-PHB,and control vectors were collected.DNA content was analyzed by flow cytometry.The proliferation level of each cell group was detected using the EdU cell proliferation assay.HepG2.2.15 cells transfected with Pcmv6-AC-GFP-PHB and the control vector were cultured in serum-free medium for 6 days.Apoptosis was measured at the indicated time points using fluorescence-activated cell sorting(FACS)-based Annexin-V/PI double staining.Results Compared with normal liver tissue,the expression of PHB in HBV-infected liver tissue was down-regulated(P<0.01).Compared with HepG2 cells,the expression of PHB in HepG2.2.15 cells was significantly decreased(P<0.01).The expression level of PHB in liver tissue after antiviral treatment(tenofovir)was significantly higher than that before treatment(P<0.01).Compared with the control vector,the proliferation rate of HepG2.2.15 cells transfected with Pcmv6-AC-GFP-PHB was significantly lower than that of the control vector,and the apoptosis rate of HepG2.2.15 cells transfected with the Pcmv6-AC-GFP-PHB vector was significantly higher than the control vector(P<0.01).Conclusion HBV down-regulates the expression of inhibin to promote the proliferation and survival of hepatocellular carcinoma cells.
作者
刘俊英
邵建营
刘洋
李涵
孔鑫
赵媛
范玉梅
吴斌
赵明
Liu Junying;Shao Jianying;Liu Yang;Li Han;Kong Xin;Zhao Yuan;Fan Yumei;Wu Bin;Zhao Ming(Zhoukou Central Hospital,Zhoukou 466000,China)
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2023年第3期288-292,共5页
Chinese Journal of Hepatology
基金
周口市科技攻关计划项目(2021GG02035、2021GG02036)
北京医卫健康公益基金会医学科学研究基金资助项目(YWJKJJHKYJJ-Q1201)。
关键词
乙型肝炎病毒
抑制素
肝细胞癌
Hepatitis B virus
Prohibitin
Hepatocellular carcinoma
