Expression analysis of genes involved in oxaliplatin-resistance in Colorectal cancer cell line

Abstract

Due to the increasing prevalence of CRC and the development of drug resistance especially to oxaliplatin, it is necessary to investigate the development of resistance to drugs used in conventional treatment methods. Therefore, in this study expression analysis of the genes involved in oxaliplatin resistance in colorectal cell line were investigated. Purpose: The aim of this project was to evaluate the expression of the genes involved in oxaliplatin resistance. Procedure: Both oxaliplatin resistant Human colorectal Caco2 and regular Caco2 cell lines were grown as monolayer in RPIM 1640 medium supplemented with 10% of heat-inactivated FBS and cultured at 37˚C in a humidified atmosphere of 5% CO2. Subsequently both cell lines were treated with different concentrations of oxaliplatin. After 48 hours, total RNA of cells was extracted and the synthesize of cDNA was completed through a reverse transcriptase. Finally, expression analysis of some of the involved genes in oxaliplatin resistance were done in all of the groups using Realtime PCR method. All the results were analysed using ΔΔCt method. Findings: The results of Real-time PCR indicate that the expression of the genes including ATP7 ،ERCC1 ،AKT1 ،mTOR ، BCL-XL ،GpX4 ،NRF2 ،Vimentin ،SNAIL1 and Twist1 after oxaliplatin treatment were increased in resistant cell line compared to the sensitive cell line, while the expression of E-cadherin was decreased after oxaliplatin treatment in resistance cell line compared to the sensitive cell line.Conclusion: According to the results of this study, these genes can be used as drug targets in the destruction of resistant cells suggesting that the use of appropriate strategies to inhibit the genes including ATP7, ERCC1, AKT1, mTOR, BCL-XL, GpX4, NRF2, Vimentin, SNAIL1 and Twist1 and the enhancement of the expression of E-cadherin may be effective to reduce the resistancy in colorectal cell line.