Study Of The Molecular Mechanism Of 5-FU-acquired Drug Resistance Of Colon Cancer HCT-8 Cells Based On Whole Transcriptome Sequencing

Colorectal cancer is a common malignant disease of digestive tract,5-fu(5-fluorouracil),as an anti-metabolic antitumor drug,is the first-line drug in the treatment of advanced colorectal cancer.The chemical tolerance of 5-fu in the course of clinical treatment severely influenced the efficacy of chemotherapy,but its underlying molecular mechanisms are unclear.As a result,To explore the molecular mechanism of 5-FU acquired drug resistance in colorectal cancer is of great significance in guiding the accurate treatment of tumors.Methods Construction of HCT-8/5-FU model of colon cancer 5-fu-acquired resistant strains in vitro by drug induction.Based on RNA-Seq,analysis of differentially expressed genes(DEGs)of colon cancer HCT-8 cells and HCT-8/5-Fu cells.Combining methylation chip and TCGA database to analyze the genes that significantly increase and significantly reduce in differentially expressed genes,Screening of drug-resistance-related genes based on related literature reports.And verifying the relationship between the change of gene expression and the acquired drug resistance of colon cancer by cell function test.Result 1)A cell model HCT-8/5-Fu with 5-FU resistance(RI=3.29)was obtained by drug-induced construction in vitro.2)A total of 453 differential genes were obtained through transcription sequencing,of which 245 were up-regulated and 208 down-regulated genes.And the gene WNT6 involved in many biological processes in tumor were screened out.3)The function of WNT6 gene was validated by cell survival rate,cell apoptosis,cell cycle and cell migration experiment.The results showed that the sensitivity of HCT-8/5-FU cells to drugs could be improved when the WNT6 gene was interferenced.And the cell apoptosis induced by 5-fu can be significantly improved,and the proliferation and migration of cells are also inhibited.Conclusion The cell function test can preliminarily determine that the WNT6 gene plays an important role in regulating drug resistance of colon cancer HCT-8 cells.This study explores the molecular mechanisms for the acquisition of colon cancer drug resistant and provides a new molecular target for accurate treatment of colon cancers.