[Effect of Cobaltous chloride on the transcription activity of modified human eNOS promoter]
1Institute of Tissue Transplantation and Immunology, Jinan University, Guangzhou 510632, China.
Aim:
To construct modified promoter of human endothelial nitric-oxide synthase(eNOS) and investigate transcriptional activity of the eNOS promoter via a hypoxia model of human umbilical vein endothelial cells-12(HUVEC-12) stimulated by Cobaltous chloride in vitro.
Methods:
The pGL2-eNOS-repSP3 vector was constructed by the SP1-replaced SP3 element with using DNA site-directed mutation of PCR. The cells were transfected with pGL2-eNOS-repSP3 vector and normal pGL2-eNOS-p vector, repsectively. Compared with another modified eNOS promoter(pGL2-eNOS-insSP1) the transcriptional activity of pGL2-eNOS-repSP3 vector was determined using a double luciferase reporter gene system by the stimulation of different concentrations of Cobaltous Chloride.
Results:
The pGL2-eNOS-repSP3 vector was constructed successfully. After the stimulation of Cobaltous Chloride, the transcriptional activities of both pGL2-eNOS-repSP3 and pGL2-eNOS-insSP1 increased as the concentration of Cobaltous Chloride increased. But there was not significantly statistical difference between them.
Conclusion:
There is no significant difference on transcriptional activities between the two modified vectors, pGL2-eNOS-repSP3 and pGL2-eNOS-insSP1.
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