pdx-1基因克隆及其在人肝癌细胞系SMMC-7721中的表达

doi:10.3969/j.issn.1004-616x.2006.02.001

癌变·畸变·突变 ›› 2006, Vol. 18 ›› Issue (2): 81-083.doi: 10.3969/j.issn.1004-616x.2006.02.001

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pdx-1基因克隆及其在人肝癌细胞系SMMC-7721中的表达

陈元晓1;李文林2;何志颖2;巴　月3;田　明1;訾晓渊2;张　彦1;胡以平2

1. 昆明医学院细胞生物学及遗传学教研室，云南　昆明　650031； 2. 第二军医大学细胞生物学教研室，上海　200433； 3. 郑州大学公共卫生学院，河南　郑州　450052

收稿日期:2005-06-06 修回日期:2005-10-25 出版日期:2006-03-30 发布日期:2006-03-30
通讯作者: 胡以平

Cloning the Gene Pancreatic Duodenal Homeobox-1 and Its Expression in SMMC-7721 Hepatoma Cells

CHEN Yuan-xiao1;LI Wen-lin2; HE Zhi-ying2;BA Yue3; TIAN Ming1;ZI Xiao-yuan2;ZHANG Yan1;HU Yi-ping2

1. Department of Cell Biology and Genetics, Kunming Medical College, Kunming 650031，Yunnan,China；2. Department of Cell Biology, College of Basic Medical Science, Second Military Medical University, Shanghai 200433,China；3. School of Public Health, Zhengzhou University, Zhengzhou 450052,Henan,China

Received:2005-06-06 Revised:2005-10-25 Online:2006-03-30 Published:2006-03-30
Contact: HU Yi-ping

摘要/Abstract

摘要： 背景与目的：克隆小鼠胰十二指肠同源框-12 (PEGFP-C1 pdx-1)基因，并构建pdx-1的表达载体。材料与方法：通过RT-PCR方法从小鼠胰腺总RNA中克隆pdx-1基因，并将该基因构建到pEGFP-C1和pcDNA3.0中获得pdx-1的表达载体。通过观察荧光和RT-PCR方法检测表达载体转染SMMC-7721后pdx-1基因在细胞中的表达。结果：克隆了883 bp的pdx-1全长cDNA，成功构建了pEGFP-C1 pdx-1和pcDNA3 pdx-1两种表达载体。pEGFP-C1 pdx-1转染SMMC-7721后，可观察到绿色荧光仅局限于细胞核中；通过RT-PCR方法在转染后的细胞中都检测到了pdx-1基因的转录。结论： pdx-1表达载体的构建为进一步研究pdx-1基因在胰腺发育和肝干细胞增殖与分化方面的作用奠定了基础。　　　　

关键词: 胰十二指肠同源框-1(pdx-1), 基因克隆, 基因表达　　

Abstract: BACKGROUND ＆ AIM: To clone the mouse pdx-1 gene and to construct its expression vector. MATERIAL AND METHODS： Mouse pdx-1 gene was amplified from mouse pancreas total RNA and subcloned into the vectors pEGFP-C1 and pcDNA3.0. The expression of pdx-1 was analyzed by RT-PCR and green fluorescence detection after transfection of the SMMC-7721 cell line. RESULTS： The expression vectors of pEGFP-C1 pdx-1 and pcDNA3 pdx-1 were constructed. After transfecting the SMMC-7721 cells with pEGFP-C1 pdx-1, green fluorescence was detected only in the nucleoli. By RT-PCR analysis, pdx-1 mRNA was detected in both pEGFP-C1 pdx-1 and pcDNA3 pdx-1 transfected cells. CONCLUSION： The expression vectors of pdx-1 gene were useful to investigate the development of the pancreas and the proliferation and differentiation of hepatic stem cells.

Key words: pancreatic duodenal homeobox-1 (pdx-1), gene cloning, gene expression

中图分类号:

Q782

陈元晓, 李文林, 何志颖, 巴　月, 田　明, 訾晓渊, 张　彦, 胡以平. pdx-1基因克隆及其在人肝癌细胞系SMMC-7721中的表达[J]. 癌变·畸变·突变, 2006, 18(2): 81-083.

CHEN Yuan-xiao, LI Wen-lin, HE Zhi-ying, BA Yue, TIAN Ming, ZI Xiao-yuan, ZHANG Yan, HU Yi-ping. Cloning the Gene Pancreatic Duodenal Homeobox-1 and Its Expression in SMMC-7721 Hepatoma Cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2006, 18(2): 81-083.

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pdx-1基因克隆及其在人肝癌细胞系SMMC-7721中的表达

Cloning the Gene Pancreatic Duodenal Homeobox-1 and Its Expression in SMMC-7721 Hepatoma Cells

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