The Study Of MiR-1254 Regulates Oxaliplatin Resistance By Targeting MEGF6 And FBLN2 In Colorectal Cancer Cells

Colorectal cancer(CRC)is one of the cancers that threaten human health.In addition to surgical resection,there are radiotherapy and chemotherapy in clinical treatment.Currently,oxaliplatin is widely used as a first-line chemotherapy drug for colorectal cancer,which has greatly reduced the mortality of colorectal cancer in the past few years..However,due to the emergence of oxaliplatin resistance in the chemotherapy of colorectal cancer,the efficay of CRC chemotherapy is is largely limited.A large number of studies have demonstrated that the abnormality of micro RNA(miRNA)is a sign of tumor production and is associated with the development of drug resistance to cancer.Objectives:1)To identify the miRNA and its target gene related to oxaliplatin resistance in CRC.2)To explore the biological function of miRNA and its target genes related to drug resistance.Experimental Methods:1)Based on HCT116 cells,the oxaliplatin resistant HCT116-R was constructed by gradually increasingly drug concentration and high concentration stimulation.2)The different miRNAs and genes in the two cell lines were analyzed by transcriptome sequencing and micro RNA sequencing,and then the miRNAs and potential target genes related to drug resistance were screened out by joint analysis.3)In order to knock down or over express related targets,synthetic miR-1254 mimics,inhibitors or interfering RNAs were transient transfection into cells.4)CCK-8 and q RT-PCR were used to detect the effect of miRNA and genes on cell resistance and proliferation.5)The effect of knockdown or overexpression related targets on apoptosis was detected by flow cytometry.6)Transwell and wound healing test experiments were used to explore the effects of knockdown or overexpression related targets on cell migration and invasion.Results:1)Overexpression of miR-1254 can significantly reduce apoptosis,enhance the resistance of HCT116 to oxaliplatin,and inhibit the expression of MEGF6 and FBLN2.2)Inhibition of miR-1254 expression can accelerate cell apoptosis,reduce the resistance of HCT116-R to oxaliplatin,and enhance the expression of MEGF6 and FBLN2.3)The high expression of miR-1254 can inhibit the migration and invasion of CRC cells,the reverse results was observed after inhibiting the expression of miR-1254.4)Knockdown of MEGF6 enhanced the resistance of CRC cells to oxaliplatin and reduced apoptosis.5)Knockdown of FBLN2 enhanced the resistance of CRC cells to oxaliplatin and inhibited cell migration and invasion.Conclusion:MiR-1254 can enhance the resistance of human CRC cells to oxaliplatin and reduce apoptosis by inhibiting the MEGF6 expression;meanwhile,miR-1254 can enhance the resistance of human CRC cells to oxaliplatin and inhibit cell migration and invasion by inhibiting the FBLN2 expression.The discovery of these molecular markers will provide some reference for the drug treatment of CRC.