环氧化酶2启动子调控促凋亡基因BAX在卵巢癌细胞系的特异性高表达  

The Specific High Expression of Apoptosis-Inducing BAX Gene Driven by Human Cyclooxygenase-2 Promoter in Ovarian Cancer Cell Line

作　　者：于爱芳[1] 辛晓燕[1] 张文红[2] 

机构地区：[1]第四军医大学附属西京医院妇产科,西安710032 [2]第四军医大学全军基因诊断技术研究所

出　　处：《中国肿瘤生物治疗杂志》2004年第2期100-104,共5页Chinese Journal of Cancer Biotherapy

摘　　要：目的:验证环氧化酶-2(COX-2)启动子能调控下游基因特异性在COX-2阳性的卵巢癌细胞系高表达;并以CMV启动子为对照,分析COX-2启动子的转录效率。方法:构建重组质粒COX-2-BAX和CMV-Lue。脂质体介导分别把质粒phPES2,CMV—Lue瞬时转染COX-2阳性的卵巢癌细胞系SKOV-3和COX-2阴性的结肠癌细胞系SW480,检测荧光素酶报告基因的表达情况。同法把质粒COX-2-BAX、pcDNA3-BAX转染SKOV-3和SW480,流式细胞仪检测细胞凋亡率。结果:成功构建重组质粒COX-2-BAX和CMV—Luc。COX-2-Lue转染SKOV3和SW480细胞24 h后报告基因活性分别为1554±86.5和53.7±10.9。CMV—Luc财法转染后,报告基因活性分别为9851.7±129.5和8831.0±167.3。COX-2-BAX转染SKOV-3和SW480细胞36 h后细胞凋亡率分别为10.4％,3.7％;CMV—BAX同法转染后,细胞凋亡率分别为21.7％,25.6％。结论:COX-2启动子可调控下游基因特异性地在COX-2阳性的卵巢癌细胞系中高表达,但转录效率明显低于CMV启动子。含COX-2启动子经合理修饰后可用于卵巢癌的基因治疗。Objective: To verify that COX-2 promoter can drive its downstream genes specifically in COX-2-positive o-varian cancer cells; Moreover, comparing with CMV promoter, analyze the transcript efficiency of COX-2 promoter. Methods: Contacting the recombinant plasmids named COX-2-BAX and CMV-Luc. After transient transfection liposome-mediated with the plasmids COX-2-Luc and CMV-Luc, respectively, the expression of Luciferase reporter gene was measured in COX-2-positive ovarian cancer cell line-SKOV3 and COX-2-negative colon cancer cell line-SW480. SKOV-3 and SW480 were transfected with COX-2-BAX and CMV-BAX in the same way, respectively. The apoptosis rates were measured through flow cytometry. Results: The recombinant plasmids named COX-2-BAX and CMV-Luc were constructed successfully. The expression efficiency of reporter gene was 1554 ± 86. 5 in SKOV3 and 53. 7 ± 10.9 in SW480 after 24 hours transfected with phPES2, 9851. 7 ± 129. 5 in SKOV3 and 8831. 0 ± 167. 3 in SW480 after 24 hours transfected with CMV-Luc in the same way. The apoptosis rate was 10.4% in SKOV3 and 3.7% in SW480 after transfected with COX-2-BAX, 21.7%in SKOV3 and 25. 6% in SW480 after 36 hours transfected with pcDNA3-BAX in the same way. Conclusions: COX-2 promoter can drive its downstream genes specifically in COX-2-positive ovarian cancer cell lines, but its expression efficiency wasmarkedly lower than CMV promoters. With proper modification, COX-2 promoter is expected to be useful in gene therapy of ovarian cancers.

关 键 词：人环氧化酶2启动子 卵巢癌 基因治疗 

分 类 号：R737.31[医药卫生—肿瘤]