<p>Regulation of <em>RUNX3</em> Expression by DNA Methylation in Prostate Cancer</p>

Abstract

To investigate the role of DNA methylation in the regulation of Runt-related transcription factor 3 (RUNX3) and the effect of such mechanism on the proliferation of prostate cancer (PCa) cells. Materials and Methods: The methylation of the RUNX3 in the promoter region in PCa cells was detected by bisulfite-sequencing PCR (BSP). Following treatment of the PCa cells with DNA methylation transferase inhibitor 5-AZA-2ʹ-deoxycytidine (AZA), the effect on methylation level and expression of RUNX3 were analyzed by qRT-PCR, Western blot, and BSP assays. Furthermore, we investigated the effect of the demethylated RUNX3 on proliferation, cell cycle and apoptosis of PCa cells using CCK-8 and flow cytometry assays. Using the DNA methylation transferase (DNMT3b) knockout or overexpression models, the relationship between DNMT3b and RUNX3 methylation was further assessed by qRT-PCR, Western blot and methylation-specific PCR (MSP). Results: The results indicated that the methylation level of RUNX3 in PCa cell lines was significantly higher than that of normal prostate epithelial (RWPE-1) cells. Furthermore, treatment with AZA not only promoted the demethylation of RUNX3 but also restored the mRNA and protein expression of RUNX3, and the reactivation of expression of the later exhibited its anti-tumor effects through regulation of the cycle progression in PCa cells. Moreover, DNMT3b could regulate the expression level of RUNX3 by altering the DNA methylation of the RUNX3 in PCa cells. Conclusion: RUNX3 is hypermethylated in a panel of PCa cell lines; inhibition of DNA methylation of RUNX3 could restore its gene expression, which could promote its anticancer effect. Thus, RUNX3 may serve as a novel putative molecular target gene for PCa therapy.

Astragaloside IV promotes cardiac remodeling after myocardial infarction by inhibiting DNMT3B-mediated Runx3 methylation via downregulating LncRNA MIRT1 expression

“…Upregulation of Runx3 can inhibit the proliferation and metastasis of gastric cancer and may serve as a new target for cancer therapy (Yu et al, 2017). Studies have reported that DNMT3B could regulate the expression level of Runx3 by altering the DNA methylation of Runx3 in prostate cancer cells involved in prostate cancer progression (Yang et al, 2020). MiR-148a may improve cell proliferation and invasion by regulating Runx3 expression through regulating DNMT3B dependent DNA methylation in gastric cancer, providing a new regimen for gastric cancer therapy (Zuo et al, 2013).…”

RUNX3 mediates keloid fibroblast proliferation through deacetylation of EZH2 by SIRT1

“…It can have a therapeutic effect on keloid by increasing the sensitivity of keloid fibroblasts to PaPDT [ 13 ]. Studies have shown that RUNX3 promoter hyper-methylation is a key factor in tumorigenesis and proliferation [ 14 , 15 ]. Meanwhile, downregulation of Zeste Homolog 2 (EZH2) can reduce the methylation level of RUNX3, and silencing EZH2 can inhibit the re-methylation of tumor cells [ 16 ].…”