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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 evaluate the specific transcriptional activity of CXCR4 and Survivin promoters in breast cancer cell lines. Methods: The CXCR4 and Survivin promoter sequences were amplified by PCR and cloned into plasmid pEGFP-1 containing the enhanced green fluorescent protein (EGFP) and plasmid pGL3-Basic containing luciferase (LUC) Breast cancer cell lines (MCF-7 MDA-MB-231 and T-47D) and mammary epithelial cells (HBL-100) were transfected with liposomes respectively. The expression of EGFP was observed under a fluorescence microscope and the luciferase activities of two promoters in breast cancer cells were determined. RESULTS: The CXCR4 and Sur-vivin promoters showed specific transcriptional activity in all 3 breast cancer cells. The CXCR4 promoter activity was higher in MDA-MB-231 than in Survivin promoter (P = 0.004), but lower in Survivin promoter than in Survivin promoter (P = 0.006), both in T-47D cells The promoter activity was comparable, P = 0.615. CONCLUSIONS: CXCR4 and Survivin promoters have strong specific transcriptional activity in breast cancer cells, and have some differences in transcriptional specificity among different breast cancer cell types.