Id1强制表达对血管内皮细胞株ECV-304增殖能力的影响  

THE EFFECT OF ID1 OVER-EXPRESSION ON THE PROLIFERATION OF ECV-304 CELLS

作　　者：陈建[1] 梁光萍[1] 罗向东[1] 杨宗城[1] 

机构地区：[1]第三军医大学附属西南医院烧伤研究所,重庆400038

出　　处：《现代预防医学》2009年第14期2702-2705,2712,共5页Modern Preventive Medicine

基　　金：国家重点基础研究发展计划"973"项目(2005CB522601);国家自然科学基金重点项目(30730093)

摘　　要：[目的]为深入研究hCASK功能,构建Id1真核表达载体并转染ECV-304细胞,筛选出Id1强制表达细胞株,观察Id1上调表达对ECV-304细胞增殖能力的影响。[方法]采用RT-PCR方法获取编码Id1的cDNA序列,并将其导入真核表达载体成功构建pIRES2–EGFP-Id1重组表达质粒。将重组质粒转染人ECV-304细胞株,经G418加压筛选,成功筛选出Id1强制表达的细胞株。用蛋白免疫印迹法鉴定转染细胞中Id1基因的表达水平。采用流式细胞技术、细胞计数的方法观察筛选出的细胞株增殖能力的变化。[结果]成功建立了Id1强制表达的ECV-304细胞株。同未转染细胞相比,Id1强制表达细胞株中G0/G1细胞比率下降约4.8%,G2M期细胞比率上升约26.3%,增殖指数明显增高,细胞增殖能力增强。[结论]Id1强制表达细胞株的成功建立和鉴定为CASK、Id1基因功能研究提供了有效手段,Id1的上调表达可导致ECV-304细胞增殖能力增强。[Objective] hCASK might be an up-stream modulator of Id1, in order to further study hCASK function, Id1 over-expression cell strain was screened out and the effect of increased Id1 expression on the proliferation of ECV-304 cells was observed. [Methods] The Id1 cDNA was obtained using RT-PCR, and was inserted into pIRES2-EGFP. The recombinated eukaryocytic expression vector pIRES2-EGFP-ldl was transfected into ECV-304 cells. The stable cell strain was screened by adding G418 into cell culture medium. The protein expression levels of Id1 gene were detected by Western Blotting. Flow cytometry and cell counting were used to measure the change of the proliferation of transfected cells. [Results] Id1 over-expression cell strain was successfully screened out. The over-expression of Id1 could lower the Go/G, phase and elevate G2M phase of the cell population. [ Conclusion] The successe in constructing Id1 over-expression cell strain offered an effective way to further study the function of CASK and Id1 genes. The increased expression of Id1 could promote the proliferation of ECV-304 cells.

关 键 词：ID1 CASK 真核细胞转染 细胞增殖 

分 类 号：R-0[医药卫生]