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目的构建了人端粒酶逆转录酶(human telomerasereserse transcription,hTERT)基因核心启动子调控的肿瘤细胞特异表达载体。方法提取人SKOV3细胞的基因组,利用PCR技术克隆hTERT基因核心启动子。并将其插入带有绿色荧光蛋白(GFP)基因的表达载体中。用脂质体转染法,将其分别转染肿瘤细胞和正常细胞,检测肿瘤细胞和正常细胞中GFP差异表达。结果HTERT基因核心启动子调控的表达载体,在所检测的3种肿瘤细胞中均具有明显的转录活性;而在正常细胞中则无明显的转录活性。结论hTRET基因核心启动子具有肿瘤特异性,构建hTERT基因核心启动子的载体可能是一种新型和有希望肿瘤治疗的途径。
Objective To construct a tumor cell specific expression vector regulated by human telomerase reverse transcriptase (hTERT) gene core promoter. Methods The genome of human SKOV3 cells was extracted and the core promoter of hTERT gene was cloned by PCR. And inserted into an expression vector carrying the green fluorescent protein (GFP) gene. Lipofectamine was used to transfect tumor cells and normal cells respectively and detect the differential expression of GFP in tumor cells and normal cells. Results The expression vector regulated by the core promoter of HTERT gene had obvious transcriptional activity in all the three kinds of tumor cells tested, but no obvious transcriptional activity in normal cells. Conclusion The core promoter of hTRET gene is tumor-specific. The construction of vector with hTERT gene core promoter may be a new and promising approach for tumor therapy.