14-3-3θ基因过表达载体的构建及其过表达乳腺癌细胞株的建立  

Vector Construction of Over Expressed Gene 14-3-3θ and Its Breast Cancer Cell Line Build

作　　者：杨继鑫[1] 王洋杨 魏洪亮[1] 陈聪[1] 杨玉庆[1] 李信[1] 李南林[1] 

机构地区：[1]第四军医大学西京医院血管内分泌外科 [2]解放军94070部队卫生所

出　　处：《现代生物医学进展》2015年第15期2820-2823,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(810721179)

摘　　要：目的:为进一步研究14-3-3θ基因对乳腺癌生物学行为的影响,构建14-3-3θ基因真核细胞过表达重组载体并建立其过表达的乳腺癌细胞株。方法:合成的14-3-3θ基因重组在p ENTR 3C DUAL载体中,课题组采用DNA重组技术将14-3-3θ基因剪切分离后,插入真核表达质粒载体pc DNA4.0中,重组得到真核过表达载体pc DNA4.0-14-3-3θ,并用其转染293T细胞后用western blot进行验证,将经过验证的真核过表达载体pc DNA4.0-14-3-3θ转染乳腺癌细胞后用zeocin筛选,筛选出的乳腺癌细胞经过western-blot和PCR验证确为稳定过表达14-3-3θ的乳腺癌细胞。结果:成功构建14-3-3θ基因过表达真核载体并建立其过表达乳腺癌细胞株,揭示14-3-3θ基因可能在乳腺癌的早期诊断和预后预测中具有重要作用。Objective： In order to further study the effect of biology behavior of the gene 14-3-3θ to breast cancer, to build a recombinant vector and breast cancer cell line which can over express gene 14-3-3θ. Methods： The gene 14-3-3θ in the pENTR 3C DUAL is synthesized by a company, we put the gene 14-3-3θ into the vector pcDNA4.0 which can product 14-3-3θ in eukaryotic cells by recombinant DNA technology and then transfect the 293T, we identify the overexpression of the recombinant vector by the western blot and PCR. At last, we transfect the breast cancer cell by the recombinant vector and screen the breast cancer cell with Zeocin. Results： Suecessfully construct the recombinant vector and breast cancer cell line which can over express 14-3-3θ. Which shows that gene 14-3-3θ may play an important role in the early diagnosis and prognosis of breast cancer prediction.

关 键 词：14-3-3θ PC DNA4.0 基因重组技术 乳腺癌 

分 类 号：Q81[生物学—生物工程]