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The Specific High Expression of Apoptosis-Inducing BAX Gene Driven by Human Cyclooxygenase-2 Promoter in Ovarian Cancer Cell Line
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Abstract:
Objective:To verify that COX-2 promoter can drive its downstream genes specifically in COX-2-positive ovarian cancer cells; Moreover, comparing with CMV promoter, analyze the transcript efficiency of COX 2 promoter. Methods:Contructing 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 ytometry.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 promoter's. With proper modification, COX-2 promoter is expected to be useful in gene therapy of ovarian cancers.
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