Establishment Of The VCR Drug-resistant Strains Of Colon Cancer Cells And The Related MiRNA Screening

Background Drug-resistant tumor cells is the important reason for the failure of cancer chemotherapy, vincristine （vincristine, VCR） is a kind of chemicals that widely used in tumor treatment. However, long-term use can make the tumor cells to VCR resistance, and tumor resistance mechanisms are still unclear. The aim of this study was to investigate the small RNA （microRNA, miRNA） related with the colon cancer drug resistance, aims to reveal the mechanisms of drug resistance andprovide the basisto tumor cells resistant reverse.Objective To establish the Vincristine drug-resistant colon cancer cell model, and screen the miRNA associated with colon cancer cells VCR resistance.Methods 1.HCT-8 colon cancer cells was cultured, and induced using different VCR gradientconcentration, establishedthe HCT-8/VCR resistant cell lines.2.The whole-genome screen strategies, HiSeq 2500 sequencing and bioinformatics methods was used, differentiated expression of miRNAs were detected and analyzed between the drug resistance HCT-8/VCR cells and non-resistant HCT-8 cells, and constructed the difference expression of miRNAs profile.3.The targeted points of the identified different miRNAs were confirmed through target gene prediction software, and the functional annotation of the gene was performed through GO.Results 1. HCT-8/VCR resistance of colon cancer cells was established using the concentration gradient, HCT-8/VCR cells grew well in the 2 × 103 ng/mLVCR.2.1651 kinds of sequencing of miRNAs were conducted, including the differences of 48 kinds of miRNAs. Compared to the drug resistance of HCT-8 colon cancer cells,24 kinds of miRNAs showed significant statistically difference （P< 0.05）, there were 17 kinds of miRNA expression increased （P< 0.05）. The expression level of Hsa- miR- 675-3 p was the highest, up to 7.497681 times, followed by hsa-miR-100-3 p, hsa miR-125-b-1-3 p, hsa-miR-82-5 p, hsa-miR-3141, chr15₁0055, hsa-miR-582-5 p, hsa-miR-582-3 p, hsa-miR-3687. Seven kinds of miRNAs expression decreasedt （P< 0.05）, the lowest was the hsa-miR-146-a-5 P（7.75199 times）, followed by hsa-miR-1247-3 P, hsa-miR-181-a-3 P, hsa-miR-1247-5 P.Conclusion Abnormal expression of miRNAs was associated with VCR resistance in colon cancer cells. The miRNA expression differences may plays a key role in colon cancer cells VCR resistance.