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| 胰十二指肠同源盒基因-1逆转录病毒表达载体的构建及其在肝干细胞中的稳定表达 |
| 金彩霞,李文林,朱吉,田棣,张南,胡以平 |
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| (宁夏医学院医学遗传学与细胞生物学教研室,银川,750004;第二军医大学基础医学部细胞生物学教研室,上海,200433) |
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| 摘要: |
| 目的:通过克隆胰十二指肠同源盒基因-1(pancreatic duodenal homeobox-1,PDX-1),构建表达PDX-1的逆转录病毒表达载体和稳定表达PDX-1的肝原始细胞系(liver epithelial progenitor cells,LEPCs),以研究肝干细胞向胰腺细胞的转分化潜能.方法:通过PCR方法克隆PDX-1基因全长,将其构建到pMSCVpuro逆转录病毒载体中获得pMSCV PDX-1 puro载体.将该载体导入Phoenix包装细胞系,进而采用乒乓转染的方法感染包装细胞PT67,获得高滴度稳定产毒的PT67细胞系.结果:成功构建了pMSCV PDX-1 puro载体,并转染PT67细胞.利用PT67细胞系产生的病毒上清可以高效的感染体外培养的肝干细胞LEPCs,获得稳定表达PDX-1基因的LEPCs(LEPCs PDX-1).结论:PDX-1逆转录病毒表达载体的成功构建和稳定表达PDX-1的肝干细胞系的获得为研究LEPCs向胰腺细胞的转分化奠定了基础. |
| 关键词: 胰十二指肠同源盒基因-1、逆转录病毒科、肝、干细胞 |
| DOI：10.3724/SP.J.1008.2006.00834 |
| 投稿时间：2006-03-07修订日期：2006-06-07 |
| 基金项目:国家自然科学基金(30270668,30200138). |
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| Construction of pancreatic duodenal homeobox-1 recombinant retroviral vector and its stable expression in liver progenitor cells |
| JIN Cai-xia,LI Wen-lin,ZHU Ji,TIAN Di,ZHANG Nan,HU Yi-ping |
| (宁夏医学院医学遗传学与细胞生物学教研室,银川,750004;第二军医大学基础医学部细胞生物学教研室,上海,200433) |
| Abstract: |
| Objective： To construct a recombinant retroviral vector expressing pancreatic duodenal homeobox-1 （PDX-1） gene and express the PDX-1 expression cassette in liver progenitor cells （LEPCs）, so as to study the conversion of hepatic cells to pancreatic-like cells, Methods： The full length of PDX-1 gene was amplified by PCR and subcloned into pMSCVpuro vector. The pMSCV PDX-1 puro was then introduced into Phoenix package cells and the cell culture supernatant was used for Ping- Pong infection of another packaging cell line PT67 to obtain stable virus-producing cell line. Results： The pMSCV PDX-1 puro vector was successfully constructed and transfected into PT67 cells. The viral supernatants of PT67 cells efficiently infected LEPCs in vitro and the infected LEPCs stably expressed PDX-1 gene, Conclusion： The pMSCV PDX-1 puro vector and PDX-1 expressing LEPCs have been successfully constructed, laying a basis for studying the transdifferentiation of liver progenitor cells to pancreas lineage cells. |
| Key words: pancreatic duodenal homeobox-1 retroviridae liver stem cells |
