The RNAi-mediated MDR1 adenovirus vector.docVIP

 PAGE \* MERGEFORMAT 19 The RNAi-mediated MDR1 adenovirus vector 【Abstract】 Objective of the RNAi-mediated MDR1 adenovirus vector of RNA interference MDR1 gene on human hepatocellular carcinoma cells. Methods According to MDR1mRNA sequence-specific shRNA vector expressing MDR1mRNA adenovirus shuttle plasmid pshuttle  MDR1. In vivo with recombinant adenovirus vector for the pAd  MDR1 transfected human hepatoma cells SMMC-7721, in order to FCM detection of cell surface membrane protein P  gp expression rate in order to confocal microscopy of intracellular Rh123 retention, Western Blot Detection of P  gp protein in the amount of change. The results build into pshuttle  MDR1 by restriction enzyme digestion and PCR confirmed that in line with the design, the pAd  MDR1 transfected hepatoma cells SMMC-7721 after, FCM detection of cell surface membrane protein P  gp expression positive rate of 22.9% and 30.8% , much lower than the control group (85.8%). Western Blot The virus-infected SMMC  7721 / R of P  gp were significantly lower than the control SMMC  7721 / R, with the SMMC  7721 / S cells close to. Conclusion successfully constructed pshuttle  MDR1 adenovirus vector, and effectively interfered with small hepatocellular carcinoma cell SMMC-7721 MDR1 expression. Keywords: hepatocellular carcinoma cells, RNAi multidrug resistance gene adenovirus vector 0 Introduction Chemotherapy is still an important means of comprehensive treatment of liver cancer is one, but the tumor multidrug resistance as an obstacle to chemotherapy. MDR1 gene and its products including over-expression is an important mechanism of multi-drug resistance [1]. We have to use RNAi technology can be significantly inhibited at the cellular level of hepatoma cells SMMC-7721 and Bel-7402 of the MDR1 expression [2,3]. To further improve the transfection efficiency of cells as well as the late animal preparation. We designed for the MDR1 construct the RNAi-mediate