The Role Of FAM64A In Prostate Cancer And Its Molecular Mechanism

Objective:1.To assay the difference of FAM64 A expression between the prostate cancer and normal tissue;2.To research the biological function and mechanism of FAM64 A in prostate cancer cells;3.To investigate the role of FAM64 A in the progression of the prostate cancer and its molecular biological mechanism.Methods:1.Using the GEO database(screening and downloading 5 public datasets,including GSE8511,GSE45016,GSE55945,GSE38241 and GSE1795)and TCGGA database assays the differences in FAM64 A expression between the prostate cancer and normal tissue,and using the database data assays the FAM64 A expression levels versus clinical staging and prognosis of the prostate cancer.2.Utilizing lentiviral-wrapped sh RNA plasmids knocks down the expression of the FAM64 A gene in DU145 and PC-3 cell lines,and using the RT-q PCR and Western-Blot techniques detect the Interference efficiency.3.The effects of the FAM64 A gene on the proliferation and apoptosis in prostate cancer cells were investigated by Cell Cytometry,Cell Counting Kit-8(CCK-8),Cell colony formation and Flow cytometry.4.DU145 cells expressing Luciferase were constructed and using living body imaging system(IVIS)analyses the effect of knocking-down of FAM64 A gene on tumor growth in nude mice.5.To identify the downstream target of FAM64 A and predict its potential function and possible molecular biological mechanism in the prostate cancer,we construct a proliferation network by using gene expression microarray technology,combining KEGG pathway and GO enrichment analysis.Results:1.Assaying GEO and TCGA data sets showed that FAM64 A was significantly overexpressed in PCA compared with normal samples.In addition,patients with high expression of FAM64 A were found to have higher clinical staging and shorter overall survival.2.The results of cell proliferation and apoptosis showed that knocking down the expression of FAM64 A in DU145 and PC-3 cells reduced the cell proliferation ability,and knocking down the expression of FAM64 A in DU145 and PC-3 cells significantly inhibited the colony formation,and inhibition of FAM64 A expression in DU145 and PC-3 cells induced apoptosis of prostate cancer cells.3.Through constructing the subcutaneous xenograft tumor model of nude mice with prostate cancer,we found that the weight and volume of nude mouse transplant tumors decreased after knocking down FAM64 A gene,indicating that knocking down FAM64 A gene inhibited the growth of prostate cancer cells in vivo.4.Through using gene expression microarray technology,KEGG and GO analysis,we find that FAM64 A plays a vital role in regulating multiple cancer-related pathways,including cellmatrix adhesion and the c AMP signaling pathway.By building the FAM64A-related PPI network,we determined that MAPK8,PRKACA,MAPK9,MX1,APP,GBP1,GNAI1,ITGA2 and ITGA6 are the key media for FAM64 A to regulate the progress of the prostate cancer.Conclusion:FAM64A is highly expressed in prostate cancer,and regulates proliferation,progression,apoptosis and other biological behaviors through a variety of cancer-related pathways,which is a key node in regulating the development of prostate and is closely related to the prognosis of prostate cancer.FAM64 A can be used as an important biomarker for prostate cancer.