Slug启动子荧光报告基因载体构建及活性检测

Construction of Slug Promoter Driven Reporter Gene Vector and Activity Analysis

目的:Slug是乳腺癌中与浸润和EMT密切相关的转录因子,研究其在人乳腺癌中的转录调控具有重要意义。方法:从人乳腺癌细胞系MDA-MB-231基因组中扩增Slug启动子,并用该启动子替换青色荧光蛋白报告基因载体pECFP-N1中的CMV启动子,获得由Slug启动子驱动的青色荧光报告载体pECFP-N1/Slug Promoter。以该载体瞬时转染MDA-MB-231细胞,共聚焦显微观察荧光蛋白表达效果。结果:Slug启动子驱动的荧光蛋白在MDA-MB-231细胞系有效表达。结论:成功构建了在转移性乳腺癌中特异性表达的Slug基因启动子驱动的荧光蛋白报告基因载体pECFP-N1/Slug Promoter。

Objective：Slug is a transcription factor that is closely related to invasion and EMT, and thus the investigation of its transcriptionregulation is important for understanding the mechanisms of breast cancer development. Method：Slug promoter was amplified from human breast cancer cell line MDA - MB -231 and was used to replace the CMV promoter in the cyan fluorescence reporter vector pECFP - N1, resulting the Slug promoter driven cyan fluorescence reporter vector pECFP - N1/Slug Promoter. Then human breast cancer cell line MDA -MB-23! was transiently transfected with pECFP- N1/Slug Promoter. The Slug promoter activity was monitored by cyan fluorescence intensity with Confocal laser - scanning microscope. Result：The Slug promoter driven fluorescence protein was effectively expressed in hu- man breast cancer cell line MDA - MB -231. Conclusion ：Metastatic breast cancer cell line MDA - MB -231 specific gene Slug promoter driven fluorescence reporter gene vector was successfully constructed.