HBV在HK-2细胞内的表达及对其转分化的影响  

Expression of HBV and its effects on the transdifferentiation in HK-2 cells

作　　者：徐慧 王义国[2] 刘长虹[2] 刘倩[2] 杜娟[3] 

机构地区：[1]山东省医学科学院,济南大学医学与生命科学学院,济南250062 [2]山东大学附属千佛山医院消化内科,济南250014 [3]山东大学附属千佛山医院中心实验室,济南250014

出　　处：《山东大学学报（医学版）》2013年第3期15-20,共6页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(NO.30901712);山东省科技攻关计划(2007GGW202854)

摘　　要：目的研究乙型肝炎病毒(HBV)在人近端肾小管上皮细胞(HK-2)的表达及对其转分化的影响。方法体外培养HK-2细胞,分为HK-2组、HK-2-PHY106组(空质粒PHY106转染组)、HK-2-PHY106-HBV组(PHY106-HBV质粒转染组)。用脂质体lipofectamineTM2000转染HK-2细胞。用ELISA法检测各组细胞培养上清液中HBsAg与HBeAg的含量。免疫细胞化学染色及Western blot印迹检测转染72 h后E-钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)的表达。RT-PCR法检测转染72 h后转化生长因子-1(TGF-β1)的mRNA。结果 HK-2-PHY106-HBV组细胞培养上清液中可检测到HBsAg和HBeAg的高表达;免疫细胞化学染色及Western blot印迹检测均显示HK-2-PHY106-HBV组E-cadherin表达显著下调(P<0.05),而α-SMA表达与其他两组相比明显上调(P<0.05);RT-PCR显示HK-2-PHY106-HBV组TGF-β1的mRNA表达上调(P<0.05)。结论 HBV可在HK-2细胞内高效复制,并能够导致HK-2转分化,其机制可能是通过上调TGF-β1来实现的。Objective To investigate the expression of HBV and effects on the transdifferentiation in HK-2 cells. Methods HK-2 cells were cultured in vitro and we devided them into the HK-2 group, the HK-2-PHY106-HBV group （HK-2 cells transfected with PHY106-HBV plasmid） and the HK-2-PHY106 group （HK-2 cells transfected with PHY106 control plasmid）. HK-2 cells were transfected by lipofectamineTM2000. Supernatant of culture was collected for examining HBsAg and HBeAg by using ELISA; The expression of E-cadherin and α-smooth muscle actin in HK-2 were assayed by immunocytochemistry and Western blot after the cells were transfected for 72h. TGF-β1 mRNA was detected by RT-PCR after the cells were transfected for 72h. Results The HBsAg and HBeAg were detected with a higher expression in the HK-2-PHY106-HBV group. The E-cadherin detected by immunocytochemistry and Western blot showed a significantly lower expression in the HK-2-PHY106-HBV group（P〈0.05）. However, the expression of α-SMA in the HK-2-PHY106-HBV group was higher than that of the other two groups（P〈0.05）. Additionally, we also found TGF-β1 mRNA expression was upregulated in the HK-2-PHY106-HBV group showed by RT-PCR（P〈0.05）. Conclusion HBV can be replicated highly efficiently in HK-2 cells and can make HK-2 cells occur transdifferentiation. The mechanism may be by upregulating TGF-β1 expression.

关 键 词：乙型肝炎病毒 肾小管上皮细胞 转染 细胞表型转化 转化生长因子-Β1 

分 类 号：R512.62[医药卫生—内科学]