Netrin-1慢病毒表达载体的构建及延缓TGF-β诱导HK-2细胞上皮-间充质转化的初步研究  

Construction of Lentivira1 Vector with Netrin-1 Gene and Attenuates EMT of HK-2 Cells that is Induced by TGF-β

作　　者：白久旭[1] 韩敬明[1] 李旭[1] 王东辉[1] 曹宁[1] 

机构地区：[1]沈阳军区总医院血液净化科,辽宁沈阳110015

出　　处：《现代生物医学进展》2012年第27期5234-5237,共4页Progress in Modern Biomedicine

基　　金：辽宁省科技攻关课题(2010225036)

摘　　要：目的:构建netrin-1基因的慢病毒表达载体,初步研究netrin-1在肾小管纤维化的作用。方法:将扩增的netrin-1表达片段克隆入FUW慢病毒表达载体,鉴定重组质粒正确性。利用HEK-293T细胞包装慢病毒,病毒感染人肾小管上皮HK-2细胞,Westernblot检测重组病毒netrin-1在真核细胞内表达表达;TGF-β刺激过表达netrin-1的HK-2细胞,利用Westernblot检测其纤维化指标的变化。结果:FUW-netrin-1慢病毒表达载体测序正确,并在感染病毒的细胞中检测出特异性条带,TGF-β刺激感染netrin-1慢病毒HK-2细胞的E-cadherin的下降水平比未感染病毒组低。结论:成功构建了netrin-1的慢病毒表达载体,发现netrin-1可能延缓肾小管内皮细胞发生纤维化。Objective： To construct a lentiviral vector expressing Netrin-1 gene, and to investigate the effect of netrin-1 on the process of renal tubule fibrosis. Methods： The gene fragment of netrin-1 was cloned into lentiviral vector FUW. Then the recombinant plasmid was confirmed by sequencing. The recombinant lentivirus was packaged in HEK-293T cells, and the expression of netrin- 1 in HK-2 cells was determined by Western blotting. The expression of fibrosis markers in HK-2 cells treated with TGF-β was investigated. Results： The recombinant plasmid FUW-netrin-1 was constructed and identified. The expression ofnetrin-1 in HK-2 cells was detected by Western blowtting. The loss of E-cadherin expression decreased significantly in the HK-2 cells infected by netrin-1 virus particle after TGF-β treatment compared with contronl group uninfected with netrin-1 virus. Conclusion： The lentivirus vector FUW-netrin-1 has been constructed successful, and netrin-1 may attenuate the progression of renal tubule fibrosis.

关 键 词：NETRIN-1 TGF-Β 肾小管上皮细胞 上皮-间充质转化 

分 类 号：Q75[生物学—分子生物学] Q78