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

Xian-Zhe Huang; Jun Huang; Wen-Zhao Li; Jun-Jie Wang; De-Ye Song; Jiang-Dong Ni

doi:10.1016/j.steroids.2019.108533

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

Steroids. 2020 Feb:154:108533. doi: 10.1016/j.steroids.2019.108533. Epub 2019 Oct 31.

Authors

Xian-Zhe Huang¹, Jun Huang¹, Wen-Zhao Li¹, Jun-Jie Wang¹, De-Ye Song¹, Jiang-Dong Ni²

Affiliations

¹ Department of Orthopedics, The Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, PR China.
² Department of Orthopedics, The Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, PR China. Electronic address: Nijiangdong001@csu.edu.cn.

PMID: 31678133
DOI: 10.1016/j.steroids.2019.108533

Abstract

Objective: To study roles oflncRNA-MALAT1 and miR-214 in steroid-induced avascular necrosis of the femoral head (SANFH).

Methods: MALAT1, miR-214 andosteogenic-relatedgenes(Runx2, ALP, andOCN)expressions were determined in SANFH tissue samples and human bone marrow stromal cells (BMSC) by RT-qPCR. BMSCs were verifiedbyflowcytometry. The ATF4 level was determined by western blotting and RT-qPCR. Osteogenesis inducedbyosteogenic 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.

Keywords: ATF4; MALAT1; Osteogenic differentiation; Steroid-induced avascular necrosis of the femoral head; miR-214.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Activating Transcription Factor 4 / genetics
Activating Transcription Factor 4 / metabolism*
Anti-Inflammatory Agents / pharmacology
Cell Differentiation / drug effects
Cell Proliferation / drug effects
Cells, Cultured
Dexamethasone / pharmacology
Dose-Response Relationship, Drug
Femur Head Necrosis / chemically induced
Femur Head Necrosis / metabolism*
Femur Head Necrosis / pathology
Humans
MicroRNAs / genetics
MicroRNAs / metabolism*
Osteogenesis / drug effects*
RNA, Long Noncoding / antagonists & inhibitors
RNA, Long Noncoding / genetics
RNA, Long Noncoding / metabolism*

Substances

ATF4 protein, human
Anti-Inflammatory Agents
MALAT1 long non-coding RNA, human
MIRN214 microRNA, human
MicroRNAs
RNA, Long Noncoding
Activating Transcription Factor 4
Dexamethasone