Effect of MYSM1 on proliferation and osteogenic differentiation of rat jaw bone marrow mesenchymal stem cells

Background Deubiquitination enzyme can regulate bone growth and development, and the deletion of Myb-like, SWIRM, and MPN domains 1 (MYSM1) gene may cause osteoporosis in the whole body. Currently, no studies have been found on the effect of MYSM1 on jaw development.

Objective To investigate the effect of MYSM1 on osteogenic differentiation of bone marrow mesenchymal stem cells derived from jaws.

Methods Jaw bone marrow mesenchymal stem cells (JBMMSCs) of 10 8-week-old Wistar rats were isolated and cultured by tissue digestion. The 3-5 generations of well-grown JBMMSCs were selected for osteogenic induction, and the changes of osteogenic markers OCN, Runx2, ALP and deubiquitination enzyme MYSM1 were detected by qPCR and Western Blot. JBMMSCs were transfected with lentivirus with empty transfection and lentivirus with MYSM1 knockdown, respectively, cells were divided into the control group and knockdown group, and qPCR was used to detect the knockdown efficiency. The effect of MYSM1 knockdown on cell proliferation was detected by CCK8 and cell clonal formation assay, and the cell migration ability was detected by cell scratch assay. Osteogenic expression of JBMMSCs transfected with lentivirus and MYSM1-knockdown JBMMSCS was detected by qPCR, ALP and alizarin red staining at 7 days after osteogenic induction.

Results After successful osteogenic induction of JBMMSCs, mRNA and protein expression levels of MYSM1 were increased (P＜0.05). CCK8 showed no difference in cell proliferation between control group and knockdown group (P＞ 0.05), the cell colony formation test and cell migration ability were significantly different between the two groups (P＞0.05). The mRNA expression levels of OCN, Runx2 and ALP of JBMMSCs in the knockdown group decreased at 7 days after osteogenic induction (P＜ 0.05), ALP and alizarin red staining were lighter than control group.

Conclusion MYSM1 knockdown does not significantly affect the proliferation and migration ability of JBMMSs, but the osteogenic differentiation ability of JBMMSCs with MYSM1 knockdown is decreased.