YY1-induced lncRNA XIST inhibits cartilage differentiation of BMSCs by binding with TAF15 to stabilizing FUT1 expression

Jian-Ying He; Min Cheng; Jia-Lian Ye; Chuan-Hua Peng; Jian Chen; Bin Luo; Xian-Yu Zhang; Qiang Fu

doi:10.1016/j.reth.2022.02.002

YY1-induced lncRNA XIST inhibits cartilage differentiation of BMSCs by binding with TAF15 to stabilizing FUT1 expression

Regen Ther. 2022 Mar 29:20:41-50. doi: 10.1016/j.reth.2022.02.002. eCollection 2022 Jun.

Authors

Jian-Ying He¹, Min Cheng², Jia-Lian Ye², Chuan-Hua Peng², Jian Chen², Bin Luo², Xian-Yu Zhang³, Qiang Fu⁴

Affiliations

¹ Orthopedics Department, JiangXi Provinvcial People's Hospital, The First Affiliated Hospital of Nanchang Medical College, Nanchang, 330006, Jiangxi Province, PR China.
² Orthopedics Department, People's Hospital of Poyang County, Shangrao, 333100, Jiangxi Province, PR China.
³ Orthopedics Department, Shangrao People's Hospital, Shangrao, 333400, Jiangxi Province, PR China.
⁴ Department of Rheumatology, JiangXi Provinvcial People's Hospital, The First Affiliated Hospital of Nanchang Medical College, Nanchang, 330006, Jiangxi Province, PR China.

Abstract

Introduction: The functional roles and mechanism of the XIST in osteoarthritis and the chondrogenic differentiation of BMSCs were clarified.

Methods: The expression levels of XIST, TAF15, FUT1 and YY1 were detected through quantitative RT-PCR. The protein expression of Sox9, ACAN, COL2A1 and FUT1 were detected by western blot and immunohistochemistry. The damage of cartilage tissue was detected by HE staining, and Safranin O-fast green. Alcian-Blue and Alizarin red S staining were performed to evaluate BMSCs chondrogenic differentiation. The relationship between XIST and TAF15, XIST and TAF15 were analyzed by RNA immunoprecipitation assay. Luciferase reporter assays and chromatin immunoprecipitation were performed to detect the interaction relationship between XIST and YY1. In addition, osteoarthritis mice were built to assess the function of XIST in vivo.

Results: The levels of XIST, TAF15 and FUT1 were upregulated in cartilage tissues from osteoarthritis patient. The level of XIST was decreased in BMSCs during chondrogenic differentiation. XIST overexpression inhibited the chondrogenic differentiation of BMSCs. Moreover, silencing of FUT1 reversed the effects of XIST overexpression on BMSCs chondrogenic differentiation. Mechanistically, in BMSCs, YY1 induced the expression of XIST in BMSCs, and XIST regulated FUT1 mRNA stability through targeting TAF15. Furthermore, silencing of XIST alleviated the symptoms of cartilage injury in OA mice.

Conclusion: Taken together, these results suggested that YY1 induced XIST was closely related to the chondrogenic differentiation of BMSCs and the progression of osteoarthritis by TAF15/FUT1 axis, and may be a new OA therapeutic target.

Keywords: ACAN, aggrecan; BMSCs; BMSCs, bone marrow-derived mesenchymal stem cells; ChIP, chromatin immunoprecipitation; Chondrogenic differentiation; FUT1, fucosyltransferase 1; H&E, hematoxylin and eosin; MSCs, mesenchymal stem cells; OA, osteoarthritis; Osteoarthritis; RIP, RNA immunoprecipitation; RT, room temperature; SOX9, sex-determining region Y (SRY)-box 9; TAF15, TATA-box-binding protein-associated factor 15; TFs, transcription factors; XIST; XIST, X inactive specific transcript; YY1; YY1, ying yang 1; lncRNAs, long noncoding RNAs; qRT-PCR, quantitative real-time polymerase chain reaction.