Engineering Research Center of Key Technology and Industrialization of Cell-Based Vaccine, Ministry of Education, Northwest Minzu University, Lanzhou 730030, China.
Gansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China.
Genes (Basel). 2024 Oct 16;15(10):1333. doi: 10.3390/genes15101333.
The Madin-Darby canine kidney (MDCK) cell line constitutes a key component of influenza vaccine production, but its dependence on adherent growth limits cell culture density and hinders vaccine yield. There is evidence that the use of gene editing techniques to inhibit cell adhesion and establish an easily suspended cell line can improve vaccine yield; however, the mechanisms underlying MDCK cell adhesion are unclear. Methods: In this study, we used transcriptomics to analyse differentially expressed mRNAs and miRNAs in adherent and suspension cultures of MDCK cells. : We found that claudin-1 (CLDN1) expression was downregulated in the suspension MDCK cells and that CLDN1 promotes MDCK cell-extracellular matrix adhesion. Additionally, microRNA (miR)-175 expression was upregulated in the suspension MDCK cells. Importantly, we demonstrated that miR-175 inhibits MDCK cell adhesion by targeting the CLDN1 3'-untranslated region (UTR). These findings contribute to a more comprehensive understanding of the regulatory mechanisms modulating cell adhesion and provide a basis for establishing suspension-adapted, genetically engineered cell lines. Our work could also facilitate the identification of targets for tumour therapy.
犬肾细胞（MDCK）系是流感疫苗生产的重要组成部分，但它对贴壁生长的依赖性限制了细胞培养密度，从而影响了疫苗产量。有证据表明，利用基因编辑技术抑制细胞黏附并建立易于悬浮的细胞系可以提高疫苗产量；然而，MDCK 细胞黏附的机制尚不清楚。方法：本研究利用转录组学分析了贴壁和悬浮培养的 MDCK 细胞中差异表达的 mRNA 和 miRNA。结果：我们发现悬浮 MDCK 细胞中 Claudin-1（CLDN1）的表达下调，并且 CLDN1 促进 MDCK 细胞与细胞外基质的黏附。此外，悬浮 MDCK 细胞中 miR-175 的表达上调。重要的是，我们证明 miR-175 通过靶向 CLDN1 的 3'-非翻译区（UTR）抑制 MDCK 细胞黏附。这些发现有助于更全面地了解调节细胞黏附的调控机制，并为建立悬浮适应的基因工程细胞系提供了基础。我们的工作还可以促进肿瘤治疗靶点的鉴定。
