人FOXC2真核表达载体的构建及稳定转染C-33A细胞系的建立  

Construction of eukaryotic expression vector pcDNA3.1(-)/FOXC2 and establishment of stable transfectant C-33A cell line

作　　者：成荣杰[1] 付艳[1] 

机构地区：[1]吉林大学第一医院妇产科,吉林长春130021

出　　处：《中国实验诊断学》2009年第10期1326-1329,共4页Chinese Journal of Laboratory Diagnosis

摘　　要：目的构建人FOXC2真核表达载体,并转染人宫颈癌C-33A细胞,建立稳定转染的细胞系。方法应用PCR方法从人FOXC2克隆中扩增其cDNA编码区序列,并将其构建于pcDNA3.1(-)真核表达载体。经PCR和测序鉴定后,利用脂质体进行宫颈癌C-33A细胞的转染,应用G418筛选出稳定表达FOXC2的细胞系,通过RT-PCR及Westernblot法检测FOXC2的表达情况。结果成功构建pcDNA3.1(-)/FOXC2真核表达载体,建立稳定转染FOXC2的C-33A细胞系,并鉴定了FOXC2目的基因在C-33A细胞中的过表达。结论FOXC2真核表达载体的成功构建为进一步研究FOXC2基因在宫颈癌中的功能奠定了良好的实验基础。Objective To construct eukaryotic expressing vector of human forkhead box-C2（FOXC2） and transfect cervical cancer C-33A cells so as to establish stable C-33A cell line expressing FOXC2.Methods The full length of hFOXC2 cDNA encoding sequence was amplified by PCR from human FOXC2 clone and was inserted into pcDNA3.1（-） to construct eukaryotic expression vector consisting of FOXC2 gene.After indentified by PCR and sequencing,we transfected the recombinant into C-33A cell by lipofectamine.FOXC2 expression was analyzed using RT-PCR and Western blot.Results The recombinant vector pcDNA3.1（-）/FOXC2 was successfully constructed and established C-33A cell line stably expressing FOXC2.The FOXC2 overexpression was detected.Conclusion The construction of the eukaryotic expression vector pcDNA3.1（-）/FOXC2 will provide solid experimental foudation for further studies on the function of FOXC2 gene in uterine cervical cancer.

关 键 词：人叉头框-C2(hFOXC2) 真核表达载体 C-33A细胞 转染 

分 类 号：R737.33[医药卫生—肿瘤] Q786[医药卫生—临床医学]