CXCR4和Survivin启动子的克隆及其在乳腺癌细胞系的转录特异性分析  

Evaluation of CXCR4 and Survivin promoters transcriptional activities in breast cancer cells

作　　者：李晓霞[1] 王宝利[2] 姚智[3] 

机构地区：[1]天津医科大学基础医学院微生物学教研室,天津300070 [2]天津医科大学代谢病医院内分泌研究所,天津300070 [3]天津医科大学基础医学院免疫学教研室,天津300070

出　　处：《中华肿瘤防治杂志》2009年第5期321-324,350,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30471962)

摘　　要：目的:评估CXCR4和Survivin启动子在乳腺癌细胞系中的特异转录活性。方法:采用PCR方法扩增CXCR4和Survivin启动子序列,将其分别克隆到含有报告基因增强绿色荧光蛋白(EGFP)的质粒载体pEGFP-1和含有荧光素酶(LUC)的质粒载体pGL3-Basic,经脂质体分别转染乳腺癌细胞系(MCF-7MDA-MB-231和T-47D)和乳腺上皮细胞(HBL-100)。荧光显微镜下观察EGFP的表达情况,并测定2个启动子在乳腺癌细胞中的荧光素酶活性。结果:CXCR4和Sur-vivin启动子在3种乳腺癌细胞中均表现特异的转录活性。CXCR4启动子在MDA-MB-231中的活性高于Survivin启动子(P=0.004);而在MCF-7细胞中活性低于Survivin启动子(P=0.006),两者在T-47D细胞中的启动子活性相当,P=0.615。结论:CXCR4和Survivin启动子在乳腺癌细胞中具有强的特异转录活性,且在不同乳腺癌细胞类型表现的转录特异性有一定差异。OBJECTIVE： To identify and evaluate the transcriptional activities of CXCR4 and Survivin promoters in breast cancer cells. METHODS： CXCR4 and Survivin gene promoters were PCR amplified and cloned into the reporter plasmid pEGFP-1 and pGL3 Basic. The constructs of pEG FP/CXCR4, pEGFP/Survivin, pGL3B/CXCR4 and pGL3B/ Survivin were transfected into breast cancer cells （MCF-7, MDA-MB-231 and T-47D） and HBIA00 respectively. The expression of EGFP and luciferase were detected to evaluate the transcriptional activities of CXCR4 and Survivin promot ers. RESULTS： CXCR4 promoter showed stronger activity than Survivin promoter in MDA-MB 231 （P： 0. 004）, while weaker than Survivin promoter in MCF-7（P=0. 006）, there was no significant difference between them in T47D, P： 0. 615. CONCLUSION： The high transcriptional activities of CXCR4 and Survivin gene promoters in breast cancer cells indicate their potential utility as novel candidates for transcriptional targeting of breast cancer.

关 键 词：乳腺肿瘤 CXCR4 SURVIVIN 启动子 转录靶向性 

分 类 号：R737.9[医药卫生—肿瘤]