METH1基因截短体的构建及其对HUVEC细胞增殖的影响  

Construction of a METH1 turncated gene and its effect on proliferation of HUVEC

作　　者：彭湃[1] 张伟[2] 鲁开化[1] 韩岩[1] 夏炜[1] 易成刚[1] 

机构地区：[1]第四军医大学西京医院整形外科,陕西西安710032 [2]第四军医大学基础部微生物学教研室

出　　处：《中国美容整形外科杂志》2007年第3期230-233,共4页Chinese Journal of Aesthetic and Plastic Surgery

基　　金：国家自然科学基金资助项目(30271346)

摘　　要：目的筛选血管生成抑制因子METH1功能编码区基因片段,为进一步研究METH1抑制增生性瘢痕的分子机制奠定基础。方法设计METH1三种基因截短体,分别构建相应的重组pcDNA3.1真核表达载体,然后将其稳定转染入HUVEC细胞,用RT-PCR、Western-Blot方法检测其在细胞中的表达,最后,通过MTT及FCM法验证其对HUVEC细胞增殖的影响。结果获得了能稳定表达的三种截短体,其中METH1-T2同METH1一样具有明显抑制HUVEC细胞增殖的作用,而METH1-T1、METH1-T3抑制细胞增殖的效果不明显。结论所获METH1-T2为功能活性区片段,可以替代METH1全长进行抑制增生性瘢痕的分子机制研究。Objective To screen active fragments of the METH1 gene to further investigate the molecular mechanism of its inhibition of hypertrophic scar. Methods Three truncated genes of METH1 were constructed, and recombined pcDNA 3.1 expression vectors containing these three DNA fragments of METH1 were oonstructed, then they were transfected into the human umbilical vein enclothelial cell （HUVEC） and the expression in HUVEC was determined by RT-PCR and Western- Blot. The effects on the proliferation of HUVEC were determined using MTT and FCM. Results Three truncated genes of METH1 with stable expression were obtained, of which METH1 - T2 was able to significantly inhibit the proliferation of HUVEC as METH1, while METH1 - T1 and METH1 - T3 had no significant effect. Conclusion METH1 - T2, as the funetionally active fragment, can substitute METH1 for the further study of the molecular mechanism of its inhibition in hypertrophic scar.

关 键 词：血管生成抑制因子METH1 增生性瘢痕 基因截短体 

分 类 号：R622[医药卫生—整形外科]