The Interference Of RNAi Expression Of MRP2 In Reversal Of Multidrug Resistance Of Vinblastine In Colorectal Cancer Cell Line

ObjectiveChemotherapy is straightly important means of combined therapy in colorectal cancer by means of operation, however multidrug resistance is a major reason of badly clinic chemotherapeutic effect, and the genesis mechanism of multidrug resistance is high expression of multidrug resistance associated protein2 and ABCC2 protein. RNA interference already has becomeed an effective tool of specific inhibition target gene expression after Fire firstly reported in 1998. The small interfere RNA expression vector targted to multidrug resistance associated protein2 was constructed on our study, and the silencing effect of the vector targeted to multidrug resistance associated protein2 was observed by transfected into colon cancer anti-drug cell HCT-8/V. That was in order to approach a possibility of RNAi in Reversal of drug Resistance of tumor and explore a new strategy to advance tumor therpy which may provide a skill and method on our next study. Methods1 The specific siRNA sequence was designed according to the MRP2 sequence. The sequence was cloned into Pgenesil-1 and then sequence analysis was performed.2 The Pgenesil-1-MRP2-siRNA expression vector was constructed by gene recombination and transfected into colon cancer cell HCT-8/Vcell by liposome. The MRP2 expression was analyzed by FQ-PCR and Western blot.3 MTT test were applied to measure the inhibition combined with vinblastine proliferation of HCT-8/V strain.Results1 The expression vector of siRNA targeted to MRP2 gene (Pgenesil-1-MRP2-siRNA) was successfully constructed.2 The result of quantitate real time PCR and western blot showed that the expression of MRP2 mRNA and protein were decreased markedly after Pgenesil-1-MRP2-siRNA trasfected using liposomal trasfection reagent (P<0.05) and the inhibitive ration was 68.6% and 75.9% .3 The result of MTT showed that the ratio of cell apoptosis of the group of RNA interference was significantly high than the control group.Conclusion1 The expression vector of siRNA targeted to MRP2 gene (Pgenesil-1-MRP2-siRNA) was successfully constructed. 2 The deliverly of siRNA directed against MRP2 was shown not only to give efficient and specific downregulation of the expression of MRP2, inhibit proliferation of HCT-8/V in vitro,but also get a fine Reversal effect.3 These results suggest that a strategy based on siRNA targeting of MRP2 may build the foundation to the clinical management of colon carcinoma.