Analysis of WNK2 regulating cytoskeleton formation in ovarian cancer cells based on phosphorylation modification sequencing

Di-san junyi daxue xuebao

HU Bin¹  LIANG Zhen²  LI Fengjie²  JIA Yongqin²  HE Simin³ 

[1] Center of Molecular Biology, School of Life Sciences of Central South University, Changsha, Hunan Province, 410031;Department of Obstetrics and Gynecology, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038;Xiangya School of Public Health, Central South University, Changsha, Hunan Province, 410028;

关键词: ovarian cancer;    wnk2;    phosphorylation modification;    vimentin;    cytoskeleton;   

DOI  :  10.16016/j.2097-0927.202110107

来源: DOAJ

【 摘 要 】

Objective To analyze the biological processes regulated by protein kinase WNK2 in ovarian cancer cells based on protein phosphorylation modified mass spectrometry. Methods Ovarian cancer cell line SKOV3 was transfected with lentiviral vector to knockdown WNK2 and the obtained cells were further screened by drug screening. Then the total protein was extracted for protein phosphorylation modification mass spectrometry analysis. Bioinformatics analysis was adopted to detect the phosphorylation changes before and after WNK2 knockdown, and differential proteins were screened out. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were employed to explore the functions of differentially phosphorylated proteins, and experiments were performed to verify the biological processes regulated by WNK2. Results Phosphorylation modified mass spectrometry indicated that WNK2 knockdown significantly decreased the phosphorylation of 745 proteins with 1 078 altered protein sites, and increased the phosphorylation of 328 proteins with 415 altered protein sites. KEGG and GO enrichment analyses showed WNK2 influences biological processes such as cytoskeleton organization, signal transduction, cell composition, and regulation of kinase activity. Phalloidin staining indicated after WNK2 knockdown resulted in broken and disordered cytoskeleton, while its overexpression promoted cytoskeleton remodeling. Conclusion WNK2 is involved in the cytoskeleton formation in ovarian cancer cells by regulating the phosphorylation of vimentin and other proteins.

【 授权许可】

Unknown