MicroRNA-141 inhibits the differentiation of bone marrow-derived mesenchymal stem cells in steroid-induced osteonecrosis via E2F3

Osteonecrosis of the femoral head (ONFH) affects the life of patients. MicroRNA-141 (miR-141) has been found associated with proliferation of bone marrow-derived mesenchymal stem cells (BMSCs). E2F transcription factor 3 (E2F3) has been identified as the target of miR-141 to regulate cell proliferation. The aim of the present study was to investigate whether miR-141 and E2F3 were involved in the osteogenic differentiation of BMSCs during ONFH. BMSCs from 4-week-old Sprague-Dawley rats were transduced with miR-141 mimic or inhibitor lentiviruses. Alkaline phosphatase staining was performed to confirm osteogenic differentiation. Reverse transcription-quantitative PCR, luciferase reporter assays and western blot analysis were also used to examine the interaction between E2F3 and miR-141 in BMSCs from the control and ONFH rats. The lentiviral transductions were carried out successfully. The mRNA expression levels of miR-141 in ONFH were upregulated, while those of E2F3 were downregulated compared with the control rat. The luciferase reporter assays indicated that miR-141 could target E2F3. miR-141 knockdown upregulated the mRNA expression levels of E2F3. In addition, osteogenic differentiation of BMSCs was inhibited following miR-141 overexpression, but increased following miR-141 knockdown, as evidenced by the results of the alkaline phosphatase staining and western blot analysis. In conclusion, miR-141 inhibits the osteogenic differentiation of BMSCs in ONFH by targeting E2F3. These two molecules may represent novel candidates to examine in order to investigate the mechanism underlying ONFH.