Role of heme oxygenase-1 in demethylating effects on SKM-1 cells induced by decitabine

• DOI: 10.4238/2015.december.22.3
• Publication Date: 2015-12-28T10:55:13Z
• Authors: R. Gao; D. Ma; P. Wang; J. Sun; J.S. Wang; Q. Fang
• Keywords: Decitabine; Demethylating agent

We evaluated the influence of heme oxygenase-1 (HO-1) gene inhibition in myelodysplastic syndrome (MDS) cell line SKM-1 on enhancement demethylating effects decitabine p15, and explored possible mechanism.DNMT1 expression cells was silenced by being transfected a constructed siRNA with liposomes.The proliferation rates after drug treatment were detected counting kit-8 assay.The apoptotic Annexin V/PI assay flow cytometry.The expressions p16, TP73, CDH1, ESR1, PDLIM4 mRNAs real-time PCR, those HO-1, DNMT1, DNMT3A, DNMT3B, HDAC, p15 proteins measured western blot.The degree methylation analyzed Heme demethylation ©FUNPEC-RP www.funpecrp.com.brGenetics Molecular Research 14 (4): 17788-17798 (2015) using methylation-specific PCR (MSP).CCK-8 showed that HO-1 inhibited; rate treated (70.91 ± 0.05%) significantly higher than control group (53.67 0.05%).Flow cytometry combination (44.25 exceeded this alone (37.70 0.05%).MSP inhibiting increased gene.As suggested blot, protein changed when changed, associated affected DNMT1 expression.Inhibited attenuated hypermethylation CDKN2B suppressing which conducive to cooperating decitabine.In conclusion, findings study provide valuable experimental evidence for targeted MDS therapy, theoretical basis further studies.