Screening Of Anti-metastatic MiRNA And Small Molecule Compounds In Colorectal Cancer And Its Molecular Mechanism

With the progress of human science and technology,the treatment of tumors is also developing fast.From surgical resection,radiotherapy,and chemotherapy,to molecular targeted therapy,immunotherapy,and traditional Chinese medicine treatment,the survival rate of cancer patients is also continuously improved.Colorectal cancer is a tumor of the digestive tract with a high incidence among human tumors,which is very harmful to the human body.This study focuses on several key points in the treatment of colorectal cancer,including anti-cancer small molecule compounds,miRNAs,and anti-cancer traditional Chinese medicine.It is expected that through this study,small molecule inhibitors effective for the treatment of colorectal cancer will be obtained,and we can further obtain the potential drug targets of colorectal cancer.Protein kinase inhibitors are an important direction in targeted therapy research.We first obtained a protein kinase KALRN related to colorectal cancer metastasis through colorectal cancer expression data,combined with driver genes and drug target information.Based on the KALRN protein structure,we obtained a small molecule compound ZINC653 87069 through pharmacophore screening.By comparing the kinase activity and molecular properties of ZINC65387069 with various tyrosine kinase inhibitors(TKIs),we found that ZINC65387069 has obvious advantages.We examined the effect of ZINC65387069 on the migration,apoptosis and cytoskeleton in colorectal cancer cells and found that ZINC65387069 can significantly inhibit the phosphorylation of BRAF protein,and it also can inhibit cell migration,destroy the cytoskeleton,and promote apoptosis.Tumor-associated fibroblasts(CAFs)are an important part of the tumor microenvironment.We obtained the CAFs content in colorectal cancer tissues by immune infiltration analysis.According to the different gene expression patterns of different CAFs contents,we screened Atractyloside,a small molecule compound associated with CAFs in colorectal cancer.Through analyzing target protein and network construction of Atractyloside,we found that FGF1,ITGB1,EDNRA,MMP9,and ITGAV may be the key targets of Atractyloside.We tested the effect of Atractyloside on migration and cytoskeleton of colorectal cancer cells and found that Atractyloside can significantly inhibit cell migration and destroy the cytoskeleton.Because some miRNAs have tumor-inhibiting properties,finding anti-tumor miRNAs is of great significance for tumor treatment.We analyzed colorectal miRNA expression profile data of colorectal cancer and screened out multiple miRNAs.By constructing a miRNA-mRNA negative regulatory network,we obtained a key tumor suppressor miRNA:miR-4469.By analyzing its mechanism,we found that miR-4469 may regulate the metastasis of colorectal cancer by affecting cell chemotaxis.We experimentally verified the expression level of miR-4469 and its effect on the proliferation and migration capacity of colorectal cancer cells and found that miR-4469 is low in both colorectal cancer tissues and colorectal cancer cells with high metastatic potential,Moreover,the expression change of miR-4469 can significantly affect the proliferation and migration of colorectal cancer cells,showing definite anti-cancer properties.Many of the existing anti-cancer small molecule compounds are derived from Chinese herbal medicine.It is of great significance to identify the effective anti-cancer components and effective targets for understanding the anti-cancer molecular mechanism of Chinese herbal medicine.We analyzed the effective anti-cancer components of Chinese herbal medicines against colorectal cancer,constructed an interaction network between anticancer components and target proteins,and obtained a key target protein HSD11B2.We found that multiple anti-cancer components can activate HSD11B2.Through the gene expression analysis,survival analysis and co-expression gene analysis of HSD11B2,we found that HSD11B2 gene expression was significantly down-regulated in colorectal cancer tissues,which was positively correlated with the overall survival time of patients and was closely related to colorectal tissue-specific genes.Upon further analysis,we found that HSD11B2 expression continued to decrease in highly differentiated,moderately differentiated,and poorly differentiated tissues of colorectal cancer.Conclusion:1.A new protein kinase inhibitor ZINC65387069 was screened out,and it was proposed that ZINC653 87069 might inhibit colorectal cancer metastasis by inhibiting protein kinase phosphorylation.2.Atractyloside,a small molecule compound related to CAFs was screened out.It was suggested that Atractyloside may inhibit colorectal cancer metastasis by targeting FGF1,ITGB1,EDNRA,MMP9,and ITGAV.3.A new tumor suppressor miRNA was screened out:miR-4469,and it was suggested that miR-4469 might regulate the metastasis of colorectal cancer by affecting cell chemotaxis.4.HSD11B2,a key target of Chinese herbal medicine in the treatment of colorectal cancer,was screened out.It is suggested that activating HSD11B2 to promote colorectal cancer tissue differentiation may be the key to the function of Chinese herbal medicine.