The Reversal Effect On MDR1 Gene-Mediated Multidrug Resistance In Human Colon Carcinoma LoVo/5-Fu Cells By RNA Interference

Objective:Colon carcinoma is one of the most commonly gastrointestinal neoplasm in our country and still increasing year by year, which badly do harm to people's health.It is of significant meaning to understand the pathogenesis and metastatic patterns of colon carcinoma.The main treatment of colon carcinoma is surgery,and adjuvant chemotherapy has a very important position in colorectal cancer, especially in advanced colorectal cancer treatment and prevention of recurrence and metastasis. However,multi-drug resistance (MDR) has become the biggest problem in the treatment of various malignant tumors, including colorectal cancer.It is the main reason in the failure of chemotherapy of several cancer.In this study, our purpose is to investigate the reversal effect on MDR1 gene-mediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells by RNA Interference.Methods:A eukaryotic expression plasmid of shRNA targeting on MDR1 was constructed and was transiently transfected into human colon carcinoma LoVo/5-Fu cells. Expression of MDR1 mRNA was detected by real-time quantitative PCR (Real-time PCR).Drug sensitivity was measure by MTT. The apoptosis of cells were determined by flow cytometry assay.Results: 1,Transfection:Plasmid vector includs genes coding green fluorescent protein (GFP) gene, and transfected cells in fluorescence microscopy can stimulate the green fluorescence.Count cell numbers under different light in the same vision,and the transfection rate turns to be 60%。2,The expression of MDR1 mRNA in LoVo/5-Fu after transfected with EASY-shRNA-MDR1:After transfected with EASY-shRNA-MDR1, the expression of MDR1 mRNA in LoVo/5-Fu cells was reduced obviously(P＜0.05)。3,Drug sensitivity after transfected with EASY-shRNA-MDR1:IC50 of 5-Fu and the resistance indexes were decreased obviously(P＜0.05), the relative reverse rate of sensitivity of LoVo/5-Fu cells to 5-Fu was 74.7%。The inhibition of the expression of MDR1 reverse MDR1 gene-mediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells. Colon neoplasms; RNA interference; Multidrug resistance gene; Apoptosis; Gene therapy4,The apoptosis of cells:The apoptotic rate increased greatly(P＜0.05).Conclusions:1,MDR1 shRNA effectively inhibited the expression of MDR1.2,The inhibition of the expression of MDR1 reverse MDR1 gene-mediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells.3,RNA interference reverse MDR1 gene-mediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells.