ACBP Inhibits The Proliferation Of Colorectal Cancer Cells Through Wnt/β-catenin Signaling Pathway

Objective: colorectal cancer is the most common malignant tumor of digestive system in China.In recent years,incidence rate and mortality rate have been increasing year by year in various regions.At present,the treatment of colorectal cancer is still dominated by surgery,but the prognosis is poor,so it is urgent to find a new treatment method.Anti cancer bioactive peptide(ACBP)is a kind of bioactive peptide.In our previous research results,ACBP not only can effectively inhibit a variety of cancers,but also can inhibit the proliferation of colorectal cancer HCT116 cells,which provides a new way for us to find a treatment for colorectal cancer.The aim of this study is to investigate the molecular mechanism of the effect of ACBP on colorectal cancer cells.Methods: 10 cases of normal colorectal tissue and 10 cases of primary colorectal cancer were collected from the Affiliated Hospital of Inner Mongolia Medical University from June to December 2018.The colorectal cancer cell lines HCT116,RKO and HT29 were selected for in vitro study.The effects of different concentrations of ACBP on the clonogenesis and proliferation of colorectal cancer cells were studied by plate cloning experiment and MTT experiment,the effects of ACBP on the migration and invasion of colorectal cancer cells were studied by scratch test and Transwell invasion test.In order to further explore the molecular mechanism of ACBP inhibiting the proliferation of colorectal cancer cells,we analyzed the RNA SEQ and bioinformatics of HCT116 cells after the action of ACBP,obtained the differential expression gene spectrum and pathway enrichment map,and analyzed the signal pathway of the action of ACBP on colorectal cancer cells.Then we performed Western blot analysis on multiple targets of Wnt pathway to detect the effect of different concentrations of ACBP on the targets.Finally,the expression ofβ-catenin,a key protein of Wnt pathway,was detected to further study the site ofACBP in colorectal cancer cell line.SPSS 19.0 statistical software was used for statistical analysis of experimental data,single factor analysis of variance was used for measurement data,and independent sample t test was used for comparison between groups(P < 0.05).Results: ACBP could affect the clonogenesis and proliferation of colorectal cancer cells,and the inhibitory effect was more obvious with the increase of dose;ACBP could inhibit the migration and invasion of colorectal cancer cells;ACBP could affect the expression of phosphorylated β-catenin,which increased in a dose-dependent manner,and exerted a dose-dependent inhibition on the expression of cyclid1,met,c-myc,multiple targets of Wnt pathway It is suggested that ACBP can inhibit the proliferation of colorectal cancer cells.Conclusion: in this study,we found that ACBP can inhibit the proliferation,migration and invasion of colorectal cancer cell lines HCT116,RKO,HT29,and the inhibition is more obvious with the increase of the concentration of ACBP.We also found that ACBP inhibited colorectal cancer cells through Wnt pathway.The expression of phosphorylated β-catenin(P-β-catenin)increased with the concentration of ACBP.After phosphorylation,β-catenin was degraded by proteasome,which reduced the accumulation of β-catenin in the cytoplasm,decreased the expression of β-catenin in the nucleus and down regulated the transcription of downstream target proteins cycd1,met and c-myc,which fully explained the inhibition of ACBP on the proliferation of colorectal cancer cells.In conclusion,our results show that ACBP can promote the degradation of β-catenin and inhibit the proliferation of colorectal cancer cells through Wnt / β-catenin signaling pathway.