| 白久旭韩敬明李旭王东辉曹宁△.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*[J].,2012,12(27):5234-5237 |
| 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 ofHK-2 Cells that is Induced by TGF-β* |
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| DOI： |
| 中文关键词: Netrin-1 TGF-β 肾小管上皮细胞 上皮- 间充质转化 |
| 英文关键词: Netrin-1 TGF-β HK-2 cell EMT |
| 基金项目:辽宁省科技攻关课题(2010225036) |
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| 摘要点击次数: 669 |
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| 中文摘要: |
| 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，Western
blot 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。 |
| 英文摘要: |
| 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 of netrin-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. |
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