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ITGB1 promotes the chondrogenic differentiation of human adipose-derived mesenchymal stem cells by activating the ERK signaling

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Abstract

Adipose-derived mesenchymal stem cell (ADSC) with a high capacity of chondrogenic differentiation was a promising candidate for cartilage defect treatment. This study’s objective is to study the roles of integrin β1 (ITGB1) in regulating ADSC chondrogenic differentiations as well as the underlying mechanisms. The identity of ADSC was confirmed by flow cytometry. ITGB1 gene was overexpressed in human ADSC (hADSC) by transfection with LV003-recombinant plasmids. Gene mRNA and protein levels were examined using quantitative RT-PCR and western blotting, respectively. Differentially expressed mRNAs and proteins were characterized by next-generation RNA sequencing and label-free quantitative proteomics, respectively. ERK signaling and AKT signaling in hADSCs were inhibited by treating with SCH772984 and GSK690693, respectively. ITGB1 gene overexpression substantially increased collagen type II alpha 1 chain (COL2A1), aggrecan (ACAN), and SRY-box transcription factor 9 (SOX9) expression but suppressed collagen type I alpha 1 chain (COL1A1) expression in hADSCs. Next-generation RNA sequencing identified a total of 246 genes differentially expressed in hADSCs by ITGB1 overexpression, such as 183 upregulated and 63 downregulated genes. Label-free proteomics characterized 34 proteins differentially expressed in ITGB1-overexpressing hADSCs. Differentially expressed genes and proteins were enriched by different biological processes such as cell adhesion and differentiation and numerous signaling pathways such as the ERK signaling pathway. ERK inhibitor treatment caused substantially enhanced chondrogenic differentiation in ITGB1-overexpressing hADSCs. ITGB1 promoted the chondrogenic differentiation of human ADSCs via the activation of the ERK signaling pathway.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was funded by the National Natural Science Foundation of China (Grant Number 81472089); and Guangdong Provincial Medical Scientific Research Foundation (Grant Number A2018299, B2019038).

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Authors and Affiliations

  1. Department of Bone and Joint Surgery, The First Affiliated Hospital, Jinan University, Guangzhou, 510632, China

    Simin Luo, Wuji Li, Wenrui Wu & Zhengang Zha

  2. Department of Endocrine, The First Affiliated Hospital, Jinan University, Guangzhou, 510632, China

    Qiping Shi

Authors
  1. Simin Luo
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  2. Qiping Shi
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  3. Wuji Li
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  4. Wenrui Wu
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  5. Zhengang Zha
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Contributions

ZZ conceived and designed the study, and critically revised the manuscript. SL performed the experiments, analyzed the data and drafted the manuscript. QS, WL and WW participated in study design, study implementation and manuscript revision. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Zhengang Zha.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Electronic supplementary material

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Supplementary file1 (XLS 17 kb). ESM_1. The detailed information of differentially expressed genes

Supplementary file2 (XLS 48 kb). ESM_2. The detailed information of differentially expressed proteins

10735_2020_9918_MOESM3_ESM.xls

Supplementary file3 (XLS 65 kb). ESM_3. The merged analysis of all mRNAs and differentially expressed proteins from two omics

10735_2020_9918_MOESM4_ESM.tif

Supplementary file4 (TIF 2521 kb). ESM_4. Regulation of AKT phosphorylation and collagen II, collagen I, aggrecan, and SOX9 protein expression in hADSCs by AKT inhibitor GSK690693. hADSCs were treated with 10 nm SCH772984 for 24 h, and protein abundances were detected using western blotting analysis, with GAPDH as the internal standard. hADSC, human adipose-derived mesenchymal stem cell; ITGB1, transforming growth factor beta 1; NC, negative control; COL2A1, collagen type II alpha 1 chain; ACAN, aggrecan; COL1A1, collagen type I alpha 1 chain; SOX9, SRY-box transcription factor 9; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; AKT, protein kinase B; p-AKT, phosphorylated AKT

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Luo, S., Shi, Q., Li, W. et al. ITGB1 promotes the chondrogenic differentiation of human adipose-derived mesenchymal stem cells by activating the ERK signaling. J Mol Hist 51, 729–739 (2020). https://doi.org/10.1007/s10735-020-09918-0

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  • DOI: https://doi.org/10.1007/s10735-020-09918-0

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