基础医学与临床 ›› 2008, Vol. 28 ›› Issue (8): 863-866.

• 研究论文 • 上一篇    下一篇

CXCR4基因表达增强K562细胞的趋化作用

陈慧菁 叶韵斌 陈淑萍 周智锋   

  1. 福建省肿瘤医院肿瘤免疫研究室 福建省肿瘤医院肿瘤免疫研究室 福建省肿瘤医院肿瘤免疫研究室 福建省肿瘤医院肿瘤免疫研究室
  • 收稿日期:2007-10-31 修回日期:1900-01-01 出版日期:2008-08-25 发布日期:2008-08-25
  • 通讯作者: 叶韵斌

CXCR4 gene expression promotes K562 cells chemataxis

Hui-jing CHEN, Yun-bin YE, Shu-ping CHEN, Zhi-feng ZHOU   

  1. Fujian Provincial Tumor Hspital,Fujian Medical University Fujian Provincial Tumor Hspital,Fujian Medical University
  • Received:2007-10-31 Revised:1900-01-01 Online:2008-08-25 Published:2008-08-25
  • Contact: Yun-bin YE,

摘要: 目的 构建高表达人CXCR4蛋白的白血病细胞株并测定CXCR4基因对其侵袭转移能力的影响。方法 从外周血淋巴细胞中提取基因组RNA,用RT-PCR扩展编码CXCR4基因片段,将CXCR4基因定向克隆到含有双启动子的真核表达载体PBudCE4.1,采用酶切和序列测定方法鉴定。将所构建的重组质粒用脂质体2000转染K562细胞,Zeocin筛选阳性克隆。流式细胞术和RT-PCR对阳性克隆进行鉴定;用Transwell板检测转染与未转染CXCR4的K562细胞对趋化因子SDF-1趋化活性。结果 成功构建编码CXCR4基因的真核表达质粒PBudCE4.1/ CXCR4,经转染入K562细胞,筛选获得能稳定高表达人CXCR4蛋白的K562/CXCR4细胞;K562/CXCR4细胞对SDF-1的趋化能力较K562细胞明显增强。结论 成功构建了转染人CXCR4基因的白血病细胞株,为进一步研究白血病细胞髓外侵润的分子机制奠定基础。

Abstract: Objective To construct the tranfected cell expressing the human CXCR4 gene and the effect on its immigration. Methods The total RNA was isolated from peripheral blood monouclear cell (PBMC),the full-length CXCR4 gene was amplified by RT-PCR and was inserted into plasmids PBudCE4.1 which have two promtor, after the identification by digestion and sequencing ,the recombinant was transfected into K562 cell by lipofectamineTM2000. After screening culture by zeocin, stable transfected K562 cell line was established, and transcription and exression of CXCR4 were identified by flow cytometry; the chemotactoc activity of K562 cell transfected and untrandfected CXCR4 was analysed by Transell plate. Results The eukaryotic expression plasmid PBudCE4.1/ CXCR4 was constructed successfully. The stable trasfected K562/CXCR4 cell lines which highly express CXCR4 was established,the chemotactic activity of K562/CXCR4 was increased significiant than K562. Conclusion The CXCR4 transfected K562 cell line was successfully established, and it can make the basis for the further research on mechanism of extramedullary infiltration in leukemia