hTERT启动子调控的p53基因膀胱癌细胞靶向性表达载体构建及意义  

Construction and expression of wild-type p53 gene inducing apoptosis driven by hTERT promoter on bladder cancer cells

作　　者：符伟军[1] 史立新[1] 洪宝发[1] 王晓雄[1] 朱晓应[1] 潘进勇[1] 高江平[1] 

机构地区：[1]解放军总医院泌尿外科,北京100853

出　　处：《军医进修学院学报》2006年第4期268-270,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：中国博士后科学基金资助项目(2004035285)

摘　　要：目的:构建人端粒酶逆转录酶基因(hTERT)启动子调控的含有治疗基因的质粒,探讨在膀胱癌细胞株中特异性靶向转录表达及其潜在的临床意义。方法:将hTERT起始转录区上游的启动子序列克隆到含有绿色荧光蛋白(GFP)基因及含野生型p53基因的质粒上,分别构建成质粒phTERT-GFP及phTERT-p53。脂质体转染法瞬时转染膀胱癌细胞株T24,应用荧光显微镜、噻唑蓝(MTT)法等方法观察在转染细胞中的差异性表达及膀胱癌细胞生长的靶向性抑制作用。结果:在端粒酶阳性的膀胱细胞中观察到hTERT启动子调控的绿色荧光蛋白的靶向性稳定表达,转染hTERT启动子调控的野生型p53基因靶向性抑制膀胱癌细胞生长,但对端粒酶阴性的正常细胞无明显影响(P<0.05)。结论:成功构建的hTERT启动子调控表达的野生型p53基因和GFP基因可在膀胱癌细胞T24中靶向性表达,hTERT启动子调控表达p53基因可靶向性抑制膀胱癌细胞生长。Objective：To construct the wild-type p53 gene vector controlled by human telomerase reverse transcriptase （hTERT） promoter and to explore the inhibitory effect of wild-type p53 gene under control of this promoter on bladder cancer cell line T24 in vitro. Methods：The enhanced green fluorescent protein （EGFP） and wild-type p53 gene were cloned into the vector carrying hTERT promoter, and recombinant vectors phTERT-EGFP and phTERT-p53 were constructed. The plasmids were transfected into bladder cancer cell line T24. The gene expression was measured by observing the expression of EGFP under the fluorescent microscopy, and the cell growth of transfected T24 cells was evaluated by MTY assay, respectively. Results： Expression of EGFP driven by hTERT gene promoter was detected in T24 cells with telomerase activity, and not detected in cells without telomerase activity. Recombinant adenovirus phTERT-p53 could selectively infect T24 cells, and the proliferation of T24 cells showed significantly inhibitory effect（ P 〈 0.05 ）. Condusion： The hTERT promoter can specifically control the target gene expression in telomerase-positive bladder cancer cells but not the normal cells, suggesting that the wild-type p53 gene under control of the hTERT promoter is a promising targeted gene therapy for bladder cancers.

关 键 词：膀胱肿瘤 基因 p53 启动子 基因疗法 

分 类 号：R737[医药卫生—肿瘤]