FAM135A Inhibits Malignancy Progression Of Colorectal Cancer Through PI3K/AKT/mTOR Pathway

Objective: In recent years,as people life level of ascension and eating habits changing,the rising incidence of colorectal cancer in the world,has become the third most common malignancy worldwide In order to prevent the occurrence and development of colorectal cancer,in addition to the appropriate changes in lifestyle and nutrition distribution,clinical examination and screening is important means for the prevention and treatment of FAM135 A is 135 th in the family of similar sequences A,A study shows in the occurrence of colorectal cancer development,FAM135 A coding protein plays an inhibitory role.In this study,q RT-PCR and Western blot were used to detect the expression level of FAM135 a in colorectal cancer.The effect of FAM135 A on the biological behavior of colorectal cancer cells was investigated by cell function experiments.By combining the results of statistics and experimental tests,this study explored the role of FAM135 a in the pathway to provide new targets and ideas for diagnosis and treatment of colorectal cancer.Methods: 1.The expression level of FAM135 A in colorectal cancer tissues and normal colorectal tissues of the same patient was detected by q RT-PCR and Western blot,and the expression level of FAM135 A in cancer tissues and clinicopathological parameters were statistically analyzed.Western blot assay was used to detect the transfection efficiency of FAM135 A after silencing or overexpression in colorectal cancer cell lines.After silencing or overexpression of FAM135 A,the invasion and migration ability of SW480 cells and HCT116 cells were detected by MTT assay.2.After transfection of FAM135 A mimics and inhibitors into SW480 cells and HCT116 cells,the expression changes of PI3 K,Akt,m TOR,E-cadherin,N-cadherin,ERK,PTEN,CDK2,Bcl2,Bax and FAK related to invasion,migration and cell proliferation pathway were detected by Western blot.Results: 1.The expression level of FAM135 A in colorectal cancer tissues and normal colorectal tissues of the same patient was detected by q RT-PCR and Western blot assay.The expression level of FAM135 A in colorectal cancer tissues was relatively lower than that in matched normal colorectal tissues.Analysis of the expression level of FAM135A and clinicopathological parameters of colorectal cancer showed that the low expression of FAM135 A in colorectal cancer tissues was negatively correlated with positive lymph node metastasis(P=0.037)and degree of differentiation of cancer cells(P =0.040).However,there was no significant correlation with age,sex,tumor location,tumor size,postoperative liver metastasis,DUCK stage.Among the six colorectal cancer cell lines of RKO,SW620,SW480,HCT116,LOVO and HT29,the expression of FAM135 A in SW480 and HCT116 cells was relatively high,so we choose SW480 cells and HCT116 cells to complete the follow-up experiments.The transient transfection of FAM135 A overexpression and silence reagent was performed on SW480 cells and HCT116 cells were transiently transfected with FAM135 A mimics and inhibitors for cytofunctional experiments.MTT assay suggested that the high expression of FAM135 A may decrease the proliferative ability of colorectal cancer cells,and the low expression of FAM135 A may improve the proliferative ability of colorectal cancer cells,but there was no significant statistical difference between them;Transwell experiment results showed that high expression of FAM135 A can reduce the invasion and migration ability of colorectal cancer cells,while low expression of FAM135 A can improve the invasion and migration ability of colorectal cancer cells.2.Western blot was used to detect proteins related to the pathway of tumor invasion,migration and cell proliferation in SW480 cells and HCT116 cells,and the results showed that: Compared with the control group,the expression levels of PI3 K,Phospho-Akt,m Tor,N-cadherin,Phospho-ERK,CDK2,and BCL2 were significantly up-regulated after silenced FAM135 A,while the total Akt and total ERK levels were not significantly changed.The expression of PTEN and E-cadherin decreased significantly,while the expression of Bax and Phospho-FAK did not change significantly.After overexpression of FAM135 A,the expression levels of PI3 K,Phospho-Akt,m TOR,N-cadherin,Phospho-ERK,CDK2,and BCL2 were significantly down-regulated,while the total AKT and total ERK levels were not significantly changed.The expression levels of PTEN and E-cadherin were significantly up-regulated.Conclusion: 1.FAM135 A inhibits the invasion and migration ability of colorectal cancer cells,and may inhibit cell proliferation.2.FAM135 A can inhibit the progression of colorectal cancer through PI3K/AKT/m TOR pathway.