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[ ABSTRACT] AIM:To determine the role of transcription factor Bach1 in the functions of human microvascular en-dothelial cells ( HMVECs ) .METHODS: Bach1 siRNA was transfected into HMVECs to knock down the expression of Bach1.In vitro endothelial cell tube formation assay in Matrigel culture was used as a surrogate assay for angiogenic poten-tial.Migration of HMVECs was determined by using Transwell chambers.Cell proliferation was measured by CCK-8 assay. Real-time PCR, Western blotting, and ELISA were employed to determine mRNA expression and protein level.Reporter as-say was performed to determine vascular endothelial growth factor ( VEGF) transcriptional activity.RESULTS:Knockdown of Bach1 expression in HMVECs led to an increase in the tube formation and increased endothelial cell migration ability, whereas it has little effect on cell proliferation.Bach1 silencing increased the mRNA and protein expression of heme oxygen-ase-1 (HO-1), and enhanced VEGF transcriptional activation, and mRNA and protein expression.CONCLUSION:Bach1 silencing increases HO-1 and VEGF expression, thus promoting the cell migration and tube formation of HMVECs, indicating that Bach1 is a repressor for angiogenesis.
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Objective To observe the effects of homocysteine on the expression of estrogen receptor alpha(ER?) mRNA and the CpG island methylation of ER? gene promotor region in rats.Methods The serum level of homocysteine in rats was measured by reversed-phase HPLC.RT-PCR and methylated specific PCR(MSP) were applied to assay mRNA expression and the methylation status in promoter CpG island of ER? gene in aorta and cultured rat aortic smooth muscle cells(RASMCs).Results The homocysteine level was increased in ove and ove+H groups(P