| BackgroundColon cancer is currently a high-incidence cancer worldwide,and its pathogenesis is not very clear.Although there are related targeted drugs for treatment,its five-year survival percentage is still less than 50%,so early diagnosis,early prevention and early treatment is particularly important.In addition,because bioinformatics integrates computer,statistics and biological knowledges,it is possible to screen and study the mechanism of colon cancer occurrence and development.PurposeWe found that the regulatory role of BUB1 and miR-495-3p in colon cancer is poorly understood.Therefore,in this study,miR-495-3p inhibited the epigenetic function of colon cancer cells,and further explored whether miR-495-3p played a regulatory role through BUB1(mitotic spindle checkpoint kinase).This study explored the role of miR495-3p in the growth of colon cancer cells and its related possible mechanisms,aiming to explore new research directions for the pathogenesis of colon cancer.MethodsFirst,BUB1 and miR-495-3p were screened using bioinformatics methods,and then qPCR was used to detect the levels of mRNA and miRNA in the colorectal cancer cells.The dual luciferase experiment was used to verify the targeting relationship between miR-495-3p and BUB1,CCK-8 was used to detect cell proliferation,cell scratches were used to detect its migration ability,the plate clone experiment was used to detect cloning ability,and flow cytometry to observe cell cycle and apoptosis,qPCR and Western blotting to detect the expression of related genes.Finally,siRNAs that silence BUB1 were used to verify the results of the miR-495-3p experiment.ResultsUsing bioinformatics methods,we screened the BUB1 and miR-495-3p,and performed qPCR verification in the cell line,and found that compared with normal intestinal epithelial cells,BUB1 was highly expressed in cancer cells and miR-495-3p was low.We found that miR-495-3p can significantly reduce the fluorescence intensity of the BUB1-WT 3’-UTR group through the dual luciferase report experiment,while the BUB1-MUT 3’-UTR group has no obvious changes.This result shows that the BUB13’-UTR is a binding domain of miR-495-3p.The overexpression of miR-495-3p inhibits the proliferation,migration,cell cloning and tumorigenesis of colon cancer cells.Meanwhile,miR-495-3p can block the cell cycle and has no effect on cell apoptosis.We also observed that when miR-495-3p targets BUB1,the expression of BUB1 is reduced,and A URKA can also be reduced,and both P21 and P2 7 are highly expressed.When BUB1 is overexpressed,AURKA will also increase(inhibition decreased),and the difference between P21 and P27 was not obvious.Similarly,when siRNAs-BUB1 are used to treat colon cell lines,siRNAs can inhibit colon cancer cell proliferation,migration and cell cloning.At the same time,siRNAs can affect the cell cycle and have no effect on cell apoptosis.ConclusionmiR-495-3p may be a potential factor for the study of the mechanism of the occurrence and development of colon cancer.It can inhibit tumor progression by binding to the 3’UTR of BUB1.This study provides new research ideas for the pathogenesis of colon cancer,and may provide a new way to expand tumor diagnosis and treatment methods. |
