Differential Proteomic Analysis of MDCK Non-tumorigenic Suspension Cell Line
LIU Geng1,2,*, PEI Meng-Yuan1,2,*, QIU Zhen-Yu1,2, TANG Tian1,2, HE Ting1,2, QIAO Zi-Lin1,2,3, WANG Jia-Min1,2,3, LIU Zhen-Bin1,2,3,**
1 Gansu Tech Innovation Center of Animal Cell, Northwest Minzu University, Lanzhou 730030, China; 2 Engineering Research Center for Key Technologies and Industrialization of Cell-Based Vaccines, Ministry of Education, Northwest Minzu University, Lanzhou 730030, China; 3 Biomedical Research Center / Key Laboratory of Bioengineering and Technology State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China
Abstract：Madin-Darby canine (Canis lupus familiaris) kidney (Madin-Darby canine kidney, MDCK) cells have the advantages of standardization and high yield as stromal cells for influenza vaccine production. However, its adherent growth has hindered the scale-up of production and its tumorigenicity has prevented the dissemination of MDCK-based vaccines. This study successfully obtained a non tumorigenic suspension cell line XF06 of MDCK through monoclonal screening and suspension domestication. A nude mouse (Mus musculous) tumor experiment was conducted using XF06 and tumorigenic suspension cell line XF04. 1×107 cells were subcutaneously injected into the back of thymus free nude mice, with 10 mice in each group; Using human (Homo sapiens) cervical cancer cell lines (Hela)(with tumorigenicity) as positive controls and human embryonic lung fibroblasts (MRC5)(without tumorigenicity) as negative controls. The results showed that the screened cell line XF06 did not form tumors in nude mice. The protein expression difference of XF04 and XF06 was compared by data-independent acquisition (DIA) proteomic technology, and the tumorigenic genes were screened, and the expression levels of genes and proteins were verified. The results shows that the gene and protein expression levels of 4 proteins (baculoviral IAP repeat containing 5 (BIRC5), fascin actin-bundling protein 1 (FSCN1), insulin like growth factor 2 mRNA binding protein 1 (IGF2BP1), TNF alpha induced protein 2 (TNFAIP2)) were significantly reduced in non-tumorigenic XF06 cells (P<0.05), which might be involved in the regulation of tumorigenic MDCK cells. This study provides reference for the screening of target genes using gene editing technology to inhibit tumorigenesis.
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