LncRNA-MALAT1 promotes osteogenic differentiation through regulating ATF4 by sponging miR-214: Implication of steroid-induced avascular necrosis of the femoral head

Objective: To study roles of lncRNA-MALAT1 and miR-214 in steroid-induced avascular necrosis of the femoral head (SANFH). Methods: MALAT1, miR-214 and osteogenic-related genes (Runx2, ALP, and OCN) expressions were determined in SANFH tissue samples and human bone marrow stromal cells (BMSC) by RT-qPCR. BMSCs were verified by flow cytometry. The ATF4 level was determined by western blotting and RT-qPCR. Osteogenesis induced by osteogenic medium (OM) in BMSCs and dexamethasone (DEX) was used to inhibit osteogenesis. The alkaline phosphatase (ALP) activity was measured and ALP staining and alizarin red staining were conducted for evaluation of osteogenic differentiation. MTT assay was used for cell proliferation. The dual luciferase reporter assay was used to confirm binding between MALAT1 and miR-214, as well as miR-214 and ATF4. Results: MALAT1 was down-regulated and miR-214 was up-regulated in SANFH tissues. DEX inhibited osteogenic differentiation of BMSC in a dose-dependent manner, leading to decreased MALAT1, increased miR-214, as well as reduced ALP activity and decreased expression of RUNX2, ALP and OCN. Either overexpression of MALAT1 or inhibition of miR-214 improved DEX-induced inhibition of BMSC osteogenic differentiation. The overexpression of miR-214 reversed the effects by MALAT1. MALAT1 directly sponged miR-214 and miR-214 directly targeted ATF4. Conclusion: MALAT1 was down-regulated, while miR-214 was elevated in SANFH tissues. MALAT1 promoted osteogenesis differentiation by sponging miR-214 to upregulate ATF4.