Effect Of KLHL24 In Colon Cancer Cells And Screening Of Its Interaction Protein

Objective: pCD513B-KLHL24 vector was constructed,which was used to investigate the effects of KLHL24 on proliferation,invasion and migration of colon cancer cell lines SW480 and LOVO,and to screen for interacting proteins of KLHL24.Method: pCD513B-KLHL24 vector was constructed.Analy ze and screen KLHL24 interacting proteins by ubiquitous proteome label-free technology.pCD513B-KLHL24,psPAX2 and PMD2.G pla smids were packaged into lentivirus.The lentivirus was transfected into SW480 cells and LOVO cells,and SW480 and LOVO cell lin es stably overexpressing KLHL24 were screened with puromycin.T he expression of KLHL24 protein was detected by western blotting,and the effects of KLHL24 on proliferation,migration and invasion of SW480 and LOVO cells were detected by MTT,clone formati on and transwell.The expression of KLHL24 in SW480 and LOV O cells was detected by immunofluorescence.Immunohistochemistry was used to detect the expression of KLHL24 protein in colon can cer tissues and adjacent tissues.Results:1.SW480 and LOVO cell lines stably overexpressing KLHL24 were successfully established.2.GO analysis showed that KLHL24 was closely related to nucleotide metabolism,protein translation and localization in biological processes,and was mainly related to cytoplasm and ribosome in cell components,and was highly close to the combination of rRNA and ribosome structural components in terms of molecular function.KEGG analysis showed that KLHL24 exerted great effect on Apoptosis,Estrogen signaling,P53 signaling pathway.Protein interactional network shows that KLHL24 may has an interaction with CDK1,KRT18,BRCA2 and other proteins.3.Stable overexpression of KLHL24 had no statistical effect on proliferation,invasion and migration of SW480 and LOVO cells.4.KLHL24 was expressed in the cytoplasm and nucleus of SW480 and LOVO cells.KLHL24 was expressed in colon cancer tissues and adjacent tissues,and the expression was not statistically different.Conclusions: 1.Stable overexpression of KLHL24 had no significant effect on proliferation,invasion and migration of SW480 and LOVO cells.2.KLHL24 may has an interaction with CDK1,KRT18,BRCA2 and other proteins.