The Study Of MiR-483-3p Involved In The Chemoprevention On Colorectal Cancer Of Black Raspberry Anthocyanins

Colorectal cancer is the third most common cancer in the world and the fourth most common cause of death in the world.The occurrence and development of colorectal cancer involves changes of multiple genes and signaling pathways.Currently,potential targets for the treatment of colon cancer is research focus.MiRNA is a small-molecule and single-stranded RNA that is commonly found in plants and animals.It acts on 3'UTR of the target gene mRNA to suppress translation or directly degrade mRNA,directly affecting the growth of the body,defense system,organ formation,cell proliferation and apoptosis,energy metabolism and other physiological processes.It has found that miRNA was involved in the development of tumor,which could be used as oncogenes or tumor suppressors.MiRNA becomes a research hotspot in tumor diagnosis,targeted therapy and prognosis.As more and more abnormal expression of miRNAs in colorectal cancer was found that miRNA could be a potential target for the treatment of colon cancer in future.Black raspberry is a natural fruit rich in anthocyanin.Previous investigation found that the black raspberry freeze-dried powder had a good preventive effect on colorectal cancer and could inhibit the development of cancer or precancerous lesions.In this study,we explore whether black raspberry anthocyanin could affect the changes of miRNAs in azoxymethane(AOM)/dextran sodium sulfate(DSS)-induced colorectal cancer mouse models and human colorectal cancer cell lines.At the same time,we study the biological role and intrinsic mechanism of miR-483-3p in colorectal cancer.Firstly,AOM/DSS-induced colorectal cancer mouse and colorectal cancer cell lines were treated with black raspberry anthocyanins.The mice colon and colorectal cancer cells were detected expression of different miRNAs by miRNA gene chip and quantitative real-time PCR.In differentially expressed miRNAs,miR-483-3p was selected to study the role in colorectal cancer.Next,The target genes and related signaling pathways of miR-483-3p were predicted by bioinformatics,and verified by Western blot and quantitative real-time PCR.Finally,we detected the effects of miR-483-3p on the biological behavior of colorectal cancer cells by using MTT,wound healing assay and colony formation assays.As the results:1.The expression of miR-483-3p in colorectal cancer mice and human colorectal cancer cells decreased after treatment with black raspberry anthocyanin,suggesting that miR-483-3p may play a role in the chemoprevention on colorectal cancer of black raspberry anthocyanins.2.We found that the 3'UTR of DKK3(Dickkopf 3)mRNA contains one miR-483-3p binding site by bioinformatics analysis.The study found that the level of miR-483 was inversely proportional to the overall survival rate of patients,The level expression of DKK3 was positively correlated with the overall survival rate of patients.Black raspberry anthocyanin reduced the expression of miR-483-3p and increased the expression of DKK3 protein.The level of DKK3 protein and mRNA was significantly increased after reducing the level of miR-483-3p in colorectal cancer cells.It is suggested that miR-483-3p may be the target gene of DKK3.3.MiR-483-3p influences the role of Wnt/?-catenin signaling pathway in colorectal cancer.In human colorectal cancer cells,Silencing of miR-483-3p inhibited the activation of Wnt/?-catenin,and decreases level of ?-catenin(the key factors of Wnt/?-catenin),c-Myc,cyclin D1 and other gene.4.MiR-483-3p influences proliferation,migration and colony formation of colorectal cancer cells in terms of both mechanism and biological behavior.The decreased expression of miR-483-3p inhibited the expression of CyclinD1,CDK4 and BCL-2 and increases the expression of BAX and E-cadherin.Silencing of miR-483-3p inhibited tumor cell proliferation proliferation,migration and colony formation.The above results indicate that miR-483-3p plays an important role in colorectal cancer by regulating DKK3-Wnt/?-catenin pathways.Black raspberry anthocyanins affect the expression of miR-483-3p.MiR-483-3p may be a therapeutic target in the future.Black raspberry anthocyanin as a new chemopreventive agent has great application potential in the prevention and treatment of colorectal cancer.