Filamin B knockdown impairs differentiation and function in mouse pre-osteoblasts via aberrant transcription and alternative splicing
Xi Wang,
Qiyu Jia,
Li Yu,
Jinyong Huang,
Xin Wang,
Lijun Zhou,
Wubulikasimu Mijiti,
Zhenzi Xie,
Shiming Dong,
Zengru Xie,
Hairong Ma
Affiliations
Xi Wang
State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Clinical Medicine Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang, China; Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China; Key Laboratory of High Incidence Disease Research in Xinjiang Medical University, Ministry of Education, Urumqi, 830011, Xinjiang, China
Qiyu Jia
Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China
Li Yu
Department of Integrated Cardiology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830000, China
Jinyong Huang
Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China
Xin Wang
Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China
Lijun Zhou
School of Public Health, Xinjiang Medical University, Urumqi, 830011 Xinjiang, China
Wubulikasimu Mijiti
Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China
Zhenzi Xie
School of Basic Medicine, Xinjiang Medical University, Urumqi, 830011 Xinjiang, China
Shiming Dong
Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China
Zengru Xie
Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China; Corresponding author.
Hairong Ma
State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Clinical Medicine Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang, China; Xinjiang Clinical Research Center for Orthopedics, Urumqi, 830011, Xinjiang, China; Key Laboratory of High Incidence Disease Research in Xinjiang Medical University, Ministry of Education, Urumqi, 830011, Xinjiang, China; Corresponding author. State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Clinical Medicine Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, Xinjiang, China.
Objective: Filamin B (FLNB) encodes an actin-binding protein that is known to function as a novel RNA-binding protein involved in cell movement and signal transduction and plays a pivotal role in bone growth. This study aimed to investigate possible FLNB function in the skeletal system by characterizing the effecs of FLNB knockdown in mouse preosteoblast cells. Methods: Stable FLNB MC3T3-E1 knockdown cells were constructed for RNA-seq and alternative splicing event (ASE) analysis of genes involved in osteoblast differentiation and function that may be regulated by FLNB. Standard transwell, MTT, ALP, qPCR, Western blot, and alizarin red staining assays were used to assess functional changes of FLNB-knockdown MC3T3-E1 cells. Results: Analysis of differentially expressed genes (DEGs) in FLNB knockdown cells revealed enrichment for genes related to osteoblast proliferation, differentiation and migration, such as ITGA10, Cebpβ, Grem1, etc. Alternative splicing (AS) analysis showed changes in the predominant mRNA isoforms of skeletal development-related genes, especially Tpx2 and Evc. Functional asslysis indicated that proliferation, migration, and differentiation were all inhibited upon FLNB knockdown in MC3T3-E1 cells compared to that in vector control cells. Conclusions: FLNB participates in regulating the transcription and AS of genes required for osteoblast development and function, consequently affecting growth and development in MC3T3-E1 cells.
