论文部分内容阅读
目的研究AP-2α的一种选择性剪切及其对MCF-7细胞生物学功能的影响。方法构建AP-2α基因表达质粒,得到pAP-2α和一个C-端编码产物完全不同于AP-2α的选择性剪切pAP-2αas表达质粒。通过荧光素酶法检测pAP-2αas和pAP-2α对含存活素(survivin)启动子核心序列的荧光素酶报告质粒pLuc-441活性影响的差异;并通过药物敏感性实验观察pAP-2αas和pAP-2α对乳腺癌细胞MCF-7药物敏感性影响的差异。结果采用荧光素酶法检测共转染pAP-2αas和pLuc-441的MCF-7细胞中survivin启动子活性,与对照pAP-2α相比,pAP-2αas也能抑制pLuc-441活性,但抑制的强度低于前者;药物敏感性实验显示,在MCF-7细胞中,转染pAP-2αas显著提高了药物敏感性,与pAP-2α产生的生物学功能近似。结论 AP-2αas作为一种体内的特异性剪切形式,具有和通常AP-2α相似但不完全相同的功能,推测这是基因表达调控的特殊方式,AP-2α对基因表达的调节在某些情况下可以不通过直接结合启动子发挥作用。
Objective To study a selective cleavage of AP-2α and its effect on the biological function of MCF-7 cells. Methods AP-2α gene expression plasmid was constructed and pAP-2α and a selective pAP-2αas expression plasmid whose C-terminal coding product was completely different from that of AP-2α were obtained. The effects of pAP-2αas and pAP-2α on luciferase reporter plasmid pLuc-441 with survivin promoter were detected by luciferase assay. The effect of pAP-2αas and pAP -2α on breast cancer cell MCF-7 drug sensitivity differences. Results The survivin promoter activity in MCF-7 cells cotransfected with pAP-2αas and pLuc-441 was detected by luciferase assay. Compared with pAP-2α, pAP-2αas also inhibited pLuc-441 activity but inhibited The drug sensitivity test showed that the transfection of pAP-2αas in MCF-7 cells significantly increased the drug sensitivity and the biological function similar to that of pAP-2α. Conclusion AP-2αas, as a specific in vivo cleavage form, has similar but not identical functions to those of AP-2α, suggesting that AP-2αas is a special way of regulating gene expression. The regulation of AP-2α gene expression in some In cases it may not work by directly binding to the promoter.