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| Study on the Functional Mechanisms of DCLK1 Regulation of Carcinogenicity in MDCK Cells |
| ZHANG Fan1,2, AO Hui-Juan1,2, HU Jia-Qi2, LI Rui1,2, MA Wei-Bin2, WANG Jun2, MA Si-Heng2, LAN Wen-Lin2, TAN Xiao2, QIAO Zi-Lin1,3, YANG Kun1,2,* |
1 Engineering Research Center of the Ministry of Education for Key Technology and Industrialization of Cell Matrix Vaccine/Gansu Technology Innovation Center of Animal Cell/Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China;
2 College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China;
3 Gansu Bioengineering Materials Engineering Research Center, Lanzhou 730030, China |
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Abstract As a crucial cellular matrix in influenza vaccine production, MDCK (Madin-Darby Canine Kidney) cells' potential tumorigenicity remains a key factor constraining vaccine safety. Double cortin-like kinase 1 (DCLK1), a tumor stem cell marker, is highly expressed in diverse malignancies and participates in malignant progression, though its role in MDCK cell tumorigenicity remains unclear. Our research group proteomic studies identified differentially expressed protein DCLK1 between high-tumorigenic and non-tumorigenic MDCK cell lines. This study established stably DCLK1-low and overexpressed MDCK cell lines using lentiviral vector-mediated gene transfection. The effects of DCLK1 on multiple biological characteristics of MDCK cells were evaluated through CCK-8 assays, scratch assay, Transwell migration, and clonation formation experiments. Results showed that DCLK1 knockdown had no significant impact on cell morphology or proliferation capacity, but significantly reduced cell migration, invasion, and low-density proliferation capabilities. Conversely, overexpression enhanced these abilities. Further investigation revealed that DCLK1 knockdown downregulated tumor-associated factors including uroplakin 3A (UPK3A), while upregulating hepatoprotein B receptor 2 (ephrin type-B receptor 2, EphB2) and ataxin-1 (ATAXN1). DCLK1 overexpression showed no significant effects on these factors. Furthermore, constructing tumor-prone MDCK cell models in BALB/c nude mice (Mus musculus) and performing staining analysis on liver tissues revealed that vascular endothelial growth factor (VEGF) and vascular cell adhesion molecule 1 (VCAM-1) expression showed no clear correlation with tumor development in MDCK cells. This suggested their tumorigenicity may not be regulated by these 2 factors. The study demonstrated that DCLK1 might participate in the tumorigenic process of MDCK cells by regulating downstream target gene expression. This study provides crucial insights into understanding the mechanisms underlying MDCK cell tumorigenesis, identify potential therapeutic targets for future research on reducing tumorigenicity, and hold significant implications for enhancing vaccine safety in MDCK cell applications.
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Received: 23 June 2025
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Corresponding Authors:
*186152592@xbmu.edu.cn
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