摘要
目的构建survivin基因启动子调控的人单纯疱疹病毒胸苷激酶(humansimplexvirus-thymidinekinase,HSV-tk)真核表达载体,检测其在人喉癌细胞中的表达。方法PCR克隆survivin基因启动子与tk基因,双酶切后分别插入pCI-neo真核表达载体中,酶切鉴定后用脂质体法转染喉癌细胞(Hep-2)和正常细胞(7702),RT-PCR法比较两种细胞tk基因的表达情况。结果成功构建了survivin启动子调控的tk基因真核表达载体pCI-neo/survivin-tk;并通过RT-PCR检测出survivin启动子调控的表达载体在喉癌细胞中有表达,而在正常细胞中无明显表达。结论survivin启动子具有肿瘤特异性,有可能解决喉癌基因治疗中的特异性杀伤问题。
Objective：To construct an eukaryotic expression vector containing HSV -tk gene under the control of survivin promoter and identify the expression of the vector in human laryngeal carcinoma cells. Methods： Survivin promotor and tk gene were amplified from plasmids by PCR, and were cloned into pCI - neo vector. After the eukaryotic expression vector was confirmed by restriction enzyme digestion, it was transfected into human laryngeal carcinoma cells（Hep -2） and human liver cells（7702） by liposome. RT -PCR was used to detect the tk gene in laryngeal carcinoma cells and normal cells. Results：Tbe eukaryotic expression vector containing HSV - tk gene under the control of survivin promoter was successfully constructed. The result of RT - PCR indicated that tk gene was expressed in laryngeal carcinoma cells ,but not in normal liver cells. Conclusion：Survivin promoter is tumorspecific and maybe useful in gene therapy of tumor.
出处
《现代肿瘤医学》
CAS
2006年第5期530-532,共3页
Journal of Modern Oncology
基金
国家自然科学基金(30371445)
陕西省社发计项目(2003K10G44)
