Epigenetic changes of RUNX۲ and DLX۵ genes in osteoblastic differentiation induced by zoledronic acid in mesenchymal stem cells

Objective: Zoledronic acid as a main treatment for osteoporosis has an important role in differentiation of mesenchymal stem cells. However, mechanism of osteoblastic differentiation induction by zoledronic acid is not well understood until now. In this research we evaluate zoledronic acid effect on methylation status of RUNX۲ and DLX۵ promoters. Materials and Methods: After isolation and expansion of hMSCs from BM, they were pulse treated with ۵μM ZA for ۳h, and incubated in osteogenic differentiation medium for ۳ weeks. DNA was extracted after first, second and third weeks of culture and also from undifferentiated MSCs. After SBS treatment, gene specific methylation analysis for RUNX۲ and DLX۵ were carried out by MSP using methylated and unmethylated primers. Results: In the genes RUNX۲ and DLX۵, M and U primers of MSP amplified promoter regions of undifferentiated and osteoblastic differentiated MSCs. Therefore, methylation status in RUNX۲ and DLX۵ promoters present incomplete methylation. Conclusion: Methyltion patterns of RUNX۲ and DLX۵ don’t change during zoledronic acid osteoblastic differentiation of MSCs. Our findings show that zoledronic acid does not induce osteoblastic differentiation via epigenetic mechanisms. Therefore, zoledronic acid can induce osteoblastic differentiation in a manner independent from DNA epigenetic changes.

کلیدواژه ها:

Mesenchymal stem cells ، Zoledronic Acid ، DNA methylation ، Osteoblastic Differentiation ، زولدرونیک اسید ، متیلاسیون DNA ، سلول های بنیادی مزانشیمی ، تمایزاستئوبلاستی