The Study Of Mechanism On KLF15 Regulating VEGF Mediated Angiogenesis

Objective: The aims of this reaserch were to study the effect of KLF15 on VEGF and angiogenesis,and to reveal the new mechanism of KLF15 regulating VEGF in angiogenesis.Method:Genomic DNA was extracted from all diabetic patients.DNA samples were randomly chosen to construct DNA pools,which were as template to amplify and sequence to screen the mutations of VEGF promoters.In addition,the assocation analysis were established between different haplotypes and pathogenesis.On the basis of above research,pGL3-Basic plasmids with different promoter regions were constructed,which were used to detect the effects of different haplotypes on promoter activity by dual-luciferase reporter system.The software of MatInspector was used to analyze the bioinformatics of different promoter sequences.After that,we used ELISA and Western Blot to detecte the expression of VEGF when overexpressed or interferenced KLF15-expressing.At last,the effects of KLF15 on VEGF-mediated angiogenesis were clarified by the results of proliferation and migration experiments.Results:A single base mutation of T> C and C> G was detected at-460 and-634 of the VEGF gene promoter region by DNA pool sequencing,and association analysis revealed that both mutations significantly affected microalbuminuria levels.Dual-luciferase reporter system detection promoter activity experiments showed that different haplotypes had a significant effect on promoter activity.Then,we relied on bio-software analysis found that-460 mutation could alter the binding of transcription factor KLF15.ELISA and Western Blot results show that KLF15 can affect the expression of VEGF.Finally,KLF15 can affect the proliferation and migration of HUVEC cells,and thus affect the angiogenesis process.Conclusion: KLF15 inhibits VEGF-mediated angiogenesis.