Effect Of TRIM59 On Proliferation,invasion And Migration Of Colorectal Cancer Cells And Its Related Mechanisms

[Background]Colorectal cancer(CRC)is a kind of digestive tract malignancies.Colorectal Cancer has the third highest incidence and fifth mortality against other malignant tumors in our country.The incidence and mortality of CRC is found increasing year by year and CRC threatens seriously people life health.The diagnosis of colorectal cancer is mainly dependent on colonoscopy,imaging study and rectal examination,however,most patients diagnosed with colorectal cancer have developed into advanced stage and may miss the best treatment time.Therefore,the early diagnosis and treatment of colorectal cancer was particularly important,there will be a very important clinical significance to find a new early diagnosis of colorectal cancer markers and their therapeutic target,so as to further improve the therapeutic effect and reduce mortality.[Objective]The aim of this study was to investigate the expression of TRIM59 in colorectal cancer and its effect on the proliferation,invasion and migration of colorectal cancer cell lines.lt will also reveal the function and molecular mechanisms of TRIM59 in colorectal cancer preliminary.[Methods]1.A total of 90 human CRC tissues and their corresponding normal colorectal mucosa were surgically acquired at The First Affliated Hospital of Nanjing Medical University.The mRNA expression levels of TRIM59 were tested by quantitative real-time PCR(qRT-PCR)for detecting the collected specimens of 90 cases of colorectal cancer and further relationship between the expression of TRIM59 and the clinicopathological factors was analysed.In addition,the expression of TRIM59 levels in CRC cell line Caco-2,SW480,HT-29,LoVo,DLD-1,HCT116 and human colorectal epithelial mucosa cell line NCM460 were detected by qRT-PCR and western blot.2.The expression of TRIM59 in colorectal cancer cell lines was down-regulated by small interfering RNA.CCK-8,plate cloning and flow cytometry were used to detect the proliferation of the CRC cells wihch were transfected with siRNA.Scratch experiments and Transwell experiments were performed to observe the effect of TRIM59 on cell invasion and migration.3.Western blot was used to detect the changes of apoptosis-related proteins and Epithelial-mesenchymal Transition(EMT)proteins before and after the TRIM59 expression was inhibited4.Western blot was used to detect the expression of PI3K-AKT pathway protein in TRIM59 interference group and control group.After using the PI3K-AKT pathway-specific inhibitor LY294002,Transwell experiment were performed to observe the changes in cell invasion and migration after TRIM59 overexpression.[Results]1.The results of qRT-PCR showed that TRIM59 was highly expressed in colorectal cancer,and Chi-square Test showed that TRIM59 expression was correlated with the following clinical and pathological data:TNM stage,lymph node metastasis,tumor invasion depth and distant metastasis.In the cell experiments,the expression level of TRIM59 in colorectal cancer cell lines SW480,HT-29,LoVo,DLD-l,HCT116 was significantly higher than that in human normal intestinal epithelium cell line NCM460,whereas Caco-2 showed no expression significant statistical differences.2.After interference with TRIM59 expression in LoVo and DLD-1 cell lines,CCK-8 and cell cloning experiments showed that inhibition of TRIM59 expression can significantly reduce the proliferation of colorectal cancer cells.Flow cytometry analysis of apoptosis results showed that interference with TRIM59 can increase the apoptosis of cell lines.Western blot results showed that the expression of Bcl-2 protein was increased and the expression of Bcl-2 was decreased in TRIM59-disturbed cell lines.3.Transwell experiments and scratch experiments showed that TRIM59 had the ability to enhance the migration and invasion of colorectal cancer cells.Western blot results showed that the expression of E-cadherin increased and the expression of Vimentin and Snail decreased.4.Western blot analysis showed that the expression of phosphorylated PI3K and phosphorylated AKT was decreased in TRIM59 interferd cell lines,while the expression of PI3K and AKT had no statistically different between the control group and the experimental group.Transwell experiments showed that overexpression of TRIM59 group compared with untreated group,cell migration,invasion increased.Addition of PI3K/AKT pathway inhibitor LY294002,the cell metastasis ability was significantly reduced.[Conclusion]TRIM59 is highly expressed in colorectal cancer and is closely related to tumor stage,distant metastasis and lymph node metastasis.Interference with TRIM59 can inhibit the proliferation and metastasis of colorectal cancer cells.TRIM59 may promote cell metastasis through the PI3K/AKT pathway.TRIM59 is expected to provide new targets and ideas for the diagnosis and treatment of colorectal cancer.