HBV在HK-2细胞内的表达及对其转分化的影响

山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (3): 15-20.

HBV在HK-2细胞内的表达及对其转分化的影响

徐慧1，王义国2，刘长虹2，刘倩2，杜娟3

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

收稿日期:2012-09-07 出版日期:2013-03-10 发布日期:2013-03-10
通讯作者: 王义国（1965- ），男，硕士，主任医师，主要从事乙型病毒性肝炎及其并发症的研究。E-mail:wangyg2145@sina.com
作者简介:徐慧（1984- ），女，硕士研究生，主要从事乙型肝炎病毒相关性肾炎的研究。E-mail:xuhui163163@163.com
基金资助:
国家自然科学基金（NO.30901712）；山东省科技攻关计划（2007GGW202854）

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

XU Hui1, WANG Yi-guo2, LIU Chang-hong2, LIU Qian2, DU Juan3

1. Shandong Academy of Medical Science，School of Medicine and Life Sciences， Jinan University， Jinan 250062， China；
2. Department of Gastroenterology, Qianfoshan Hospital Affiliated to Shandong University， Jinan 250014， China；
3.Central Laboratory， Qianfoshan Hospital Affiliated to Shandong University， Jinan 250014， China

Received:2012-09-07 Online:2013-03-10 Published:2013-03-10

摘要/Abstract

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72h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72h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。

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

Abstract:

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.

Key words: Hepatitis B virus； Tubular epithelial cell；Transfection；Phenotypic transformation；Transforming growth factor-β1

中图分类号:

R512.6+2

徐慧1，王义国2，刘长虹2，刘倩2，杜娟3. HBV在HK-2细胞内的表达及对其转分化的影响[J]. 山东大学学报(医学版), 2013, 51(3): 15-20.

XU Hui1, WANG Yi-guo2, LIU Chang-hong2, LIU Qian2, DU Juan3. Expression of HBV and its effects on the transdifferentiation in HK-2 cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(3): 15-20.

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