Discovery Of A Casein Kinase Fam20C Inhibitor Of With Antitumor Activities In Vitro

Triple negative breast cancer(TNBC)is a type of breast cancer with worse prognosis,which is more invasive and metastatic than other subtypes.Fam20 C is a key factor in the migration of triple negative breast cancer cells.This article aimed to investigate the feasibility of Fam20 C as a potential target for TNBC therapy and to find Fam20 C inhibitors as a new approach for anti-TNBC therapy.The Fam20 C protein interaction network was constructed by collecting protein interaction data from the PrePPI protein interaction database,combining the GO(Gene ontology)annotation,the mechanism of Fam20 C in TNBC was studied by means of systematic biology.The data from the two cancer databases of TCGA and METABRIC were used to investigate whether there were significant differences in the mRNA expression of Fam20 C between different subtypes of breast cancer.The binding sites for selective inhibitors were determined by homologous modeling and protein crystal stacking.And the Fam20 C inhibitors were obtained by application of molecular docking,MTT cytotoxicity assays and molecular dynamics simulations.The antitumor mechanism of the inhibitor was studied by fluorescence microscopy,western blot,flow cytometry and scratch wound-healing assay.Network analysis showed that the total network of Fam20 C protein interaction consisted of 672 nodes and 974 interactions,and the function of Fam20 C in cancer was preliminarily elucidated by the combination of GO function.Transcriptional analysis showed that the mRNA expression of Fam20 C in the TCGA database was higher in the TNBC sample group than in the Non-TNBC sample group(Wilcoxon rank sum test P = 6.79E-16),consistent with the METABRIC database(Wilcoxon rank sum test P = 2.77E-19).MTT cytotoxicity test showed that FL-1607 had a certain antitumor activity against MDA-MB-468 cells and induced the morphological changes of apoptotic cells(chromatin condensation).Flow cytometry showed that the apoptotic rate increased with the dose of FL-1607 in a dose-dependent manner.The expression of Bax,Cyto.C,Caspase3 protein was up-regulated,while the expression of Bcl-2 protein was down-regulated,indicating that FL-1607 could induce the cell apoptosis via mitochondrial pathway.Scratch wound-healing assay showed that compared with the blank group,the distance of the MDA-MB-468 cells that had migrated for the scratch line of FL-1607 treatment group was shorter.In summary,Fam20 C is an effective target for the treatment of triple-negative breast cancer.FL-1607 can induce apoptosis of TNBC cell line(MDA-MB-468)and potentially inhibit cell migration.