2. Academic Validation
  3. Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling

Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling

  • Int J Mol Sci. 2025 Jan 4;26(1):395. doi: 10.3390/ijms26010395.
Rui Li 1 2 3 4 5,  Fan Zhang 1 2 3 4 5,  Lijin Wang 5,  Siya Wang 1 2 3 4 5,  Manlin Zhou 1 2 3 4 5,  Jun Wang 5,  Yiyang Zhang 1 2 3 4 5,  Xiao Tan 5,  Weiji Chen 5,  Kun Yang 1 2 3 4 5,  Zilin Qiao 1 2 3 4 5
Affiliations

Affiliations

  • 1 Engineering Research Center of Key Technology and Industrialization of Cell-Based Vaccine, Ministry of Education, Lanzhou 730030, China.
  • 2 Gansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China.
  • 3 Key Laboratory of Biotechnology & Bioengineering of State Ethnic Affairs Commission, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China.
  • 4 China-Malaysia National Joint Laboratory, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China.
  • 5 School of Life Sciences and Engineering, Northwest Minzu University, Lanzhou 730030, China.
PMID: 39796249 DOI: 10.3390/ijms26010395
Abstract

Madin-Darby Canine Kidney (MDCK) cells are a key cell line for influenza vaccine production, due to their high viral yield and low mutation resistance. In our laboratory, we established a tertiary cell bank (called M60) using a standard MDCK cell line imported from American Type Culture Collection (ATCC) in the USA. Due to their controversial tumourigenicity, we domesticated non-tumourigenic MDCK cells (named CL23) for influenza vaccine production via monoclonal screening in the early stage of this study, and the screened CL23 cells were characterised based on their low proliferative capacity, which had certain limitations in terms of expanding their production during cell resuscitation. It was thus our objective to enhance the proliferation efficiency of MDCK cells for influenza vaccine production following cell resuscitation, with a view to improving the production of non-tumourigenic MDCK cells for vaccines and enhancing the production of Influenza Virus lysate vaccines from MDCK cells through genetic intervention. We concentrated on the protein thrombosponin-1 (THBS1), which was markedly differentiated in the proteomics data of the two MDCK cells. By integrating this finding with related studies, we were able to ascertain that THBS1 exerts a significant influence on the level of cell proliferation and Apoptosis. Consequently, our objective was to investigate the impact of THBS1 expression on MDCK cell Apoptosis by verifying the differences in THBS1 expression between the two MDCK cells and by interfering with THBS1 expression in the MDCK cells. We found that the knockdown of THBS1 significantly increased the proliferation and Apoptosis of CL23 cells without causing significant changes in cell migration and invasion, and its overexpression significantly decreased the proliferation of M60 cells and increased cell migration, invasion, and Apoptosis. In addition, the TGF-β/Smad pathway target genes transforming growth factor-β1 (TGF-β1), mothers against decapentaplegic homolog 2 (SMAD2), and mothers against decapentaplegic homolog 3 (SMAD3), were significantly down-regulated in CL23 cells after THBS1 knockdown and up-regulated in M60 cells after overexpression, with consistent expression identified at both the mRNA and protein levels. The treatment of cells with TGF-β activators and inhibitors further demonstrated that THBS1 regulated MDCK cell proliferation and Apoptosis through the TGF-β/Smad signalling pathway. Finally, we found that THBS1 also regulated H1N1 Influenza Virus replication. These findings enable a comprehensive understanding of the regulatory mechanisms of THBS1 regarding MDCK cell proliferation and Apoptosis functions and the effects of Influenza Virus replication.

Keywords

MDCK cells; TGF-β pathway; THBS1; apoptosis; proliferation.

Figures
Products
Inhibitors & Agonists
Other Products