Androgen Receptor Promotes The Progression Of Gastric Cancer By Cell Cycle-related Kinase

Objective Some studies have found that AR is an independent adverse prognostic factor for patients with gastric cancer,but there is no research on the molecular mechanism of AR in gastric cancer.AR can promote the proliferation of tumor cells by promoting the expression of CCRK in liver cancer.Therefore,AR-CCRK pathway may also exist in gastric cancer to stimulate the occurrence of tumor.In this study,we will study the expression of AR and CCRK in gastric cancer.We will study the influence of AR and CCRK on gastric cancer cells,such as the role of AR and CCRK in cell migration,invasion and proliferation.We will study the regulatory relationship between AR and CCRK in gastric cancer,further explore the downstream signaling pathway regulated by AR and CCRK,and elucidate the molecular mechanism of its promoting effect.Methods In this study,we used 7 gastric cancer cells(AGS,SGC7901,MKN45,MKN28,HGC27,BGC823,and MGC803)and 1 normal gastric mucosal cell(GES-1).We constructed AR and CCRK overexpressed gastric cancer cells and knock down AR and CCRK gastric cancer cells by lentivirus transfection.(1)The expression levels of AR and CCRK in gastric cancer cells and normal gastric mucosal cells,as well as the over-expression of AR,and the expression levels of CCRK,p-gsk3,p-toy-catenin and EGFR in knocked down AR gastric cancer cells were detected by q PCR and Western blot.(2)Transwell assay was used to detect the influence of AR and CCRK on the migration and invasion of gastric cancer cells.(3)The effect of AR and CCRK on proliferation of gastric cancer cells was detected by plate cloning formation experiment.(4)The correlation between AR and CCRK gene expression levels in 591 samples of patients with gastric cancer from TCGA database was analyzed.(5)The interaction between AR protein and CCRK DNA was detected by Ch IP test.Results(1)AR m RNA was highly expressed in gastric cancer cells,and AR protein was highly expressed in gastric cancer cells(except AGS gastric cancer cells).(2)CCRK m RNA was highly expressed in gastric cancer cells,and CCRK protein was highly expressed in gastric cancer cells.(3)Transwell assay showed that the migration and invasion abilities of cells were significantly enhanced in the AR overexpressed gastric cancer cells,while the migration and invasion abilities of cells were weakened in the gastric cancer cells that were knocked down AR.In the plate colony formation experiment,the results showed that the proliferation of AR overexpressed gastric cancer cells was enhanced,and that of gastric cancer cells that were knocked down AR was weakened.(4)Transwell assay showed that in the CCRK overexpressed gastric cancer cells,the migration and invasion abilities of cells were significantly enhanced,while in the gastric cancer cells that were knocked down CCRK,the migration and invasion abilities of cells were weakened.In the plate colony formation experiment,the results showed that the proliferation of CCRK overexpressed gastric cancer cells was enhanced,and that of gastric cancer cells that were knocked down CCRKwas weakened.(5)We found a positive correlation between the expression of AR and CCRK.In Ch IP experiments,AR specifically binds to CCRK promoter in gastric cancer cells.(6)The expression levels of CCRK,p-GSK3β,p-β-catenin and EGFR were increased in AR overexpressed gastric cancer cells,while the expression levels of CCRK,p-GSK3β,p-β-catenin and EGFR in gastric cancer cells that were knocked down AR were decreased.Conclusions To sum up,we found the high expression of AR and CCRK in gastric cancer.The expression of AR can stimulate the expression of CCRK in the gastric cancer.The expression of CCRK can promote the phosphorylation of GSK3β and β-catenin,the expression of EGFR,the migration,invasion and proliferation abilities of gastric cancer cells,so as to promote the occurrence and development of gastric cancer.