Bcl‑xL mutant promotes cartilage differentiation of BMSCs by upregulating TGF‑β/BMP expression levels

Xiao,Kai; Yang,Lin; Xie,Wei; Gao,Xinfeng; Huang,Ruokun; Xie,Ming

doi:10.3892/etm.2021.10168

Bcl‑xL mutant promotes cartilage differentiation of BMSCs by upregulating TGF‑β/BMP expression levels

Authors:
- Kai Xiao
- Lin Yang
- Wei Xie
- Xinfeng Gao
- Ruokun Huang
- Ming Xie
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Affiliations: Foot and Ankle Surgery, Wuhan Fourth Hospital, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430033, P.R. China, Department of Allergy, Tongji Hospital of Tongji Medical College of HUST, Wuhan, Hubei 430033, P.R. China
Published online on: May 9, 2021 https://doi.org/10.3892/etm.2021.10168
Article Number: 736
Copyright: © Xiao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Bcl‑xL is a transmembrane molecule in the mitochondria, with apoptosis‑related and pro‑metabolic functions, that also plays a role in chondrogenesis and differentiation. A Bcl‑xL mutant, in which the GRI sequence is replaced by ELN, has no anti‑apoptotic effect, while other biological functions of this mutant remain unchanged. The present study investigated the impact of this Bcl‑xL mutant on cartilage differentiation and the expression levels of TGF‑β and bone morphogenetic protein (BMP). Human bone marrow mesenchymal stem cells (BMSCs) were transfected with Bcl‑xL and Bcl‑xL mutant (∆Bcl‑xL) overexpression vectors. The cells were divided into four groups: Control (not subjected to any transfection), EV (empty pcDNA3.1‑Bcl‑xL vector), OV (Bcl‑xL overexpression) and ∆OV (∆Bcl‑xL overexpression). Saffron and toluidine blue staining was performed to observe cartilage tissue formation. Flow cytometry was conducted to measure BMSC apoptosis. The expression levels of TGF‑β and BMP were evaluated using reverse transcription‑quantitative PCR (RT‑qPCR) and western blotting. Compared with that in the control group, the expression levels of Bcl‑xL in the OV group increased significantly (P<0.05). Western blotting and RT‑qPCR results revealed that OV and ∆OV treatment increased the expression levels of TGF‑β and BMP in transfected cells, compared to their expression in the control and EV groups (P<0.05). Saffron and toluidine blue staining results showed that cartilage formation was increased in the ∆OV and ∆OV + Bax‑/Bak‑groups to similar degrees. Cell apoptosis in the ∆OV group did not change compared with that in the control group. The Bcl‑xL mutant promoted cartilage differentiation of BMSCs and upregulated TGF‑β/BMP expression. This enhancement of chondrogenic differentiation was not related to the expression of Bax and Bak. Taken together, these findings provided for improved application of bone tissue engineering technology in the treatment of articular cartilage defects.

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