Chinese Clinical Oncology

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The experimental study of AFP-mediated lentivirus in human hepatocellular carcinoma bearing nude mice

WANG Xuan,WANG Xin,YUNA Zhenhua,XIA Bingxiang,ZHANG Yewei   

  1. Department of General Surgery,Affiliated Tumor Hospital of Nanjing Medical University, Nanjing 210009,China
  • Received:2015-07-14 Revised:2015-09-08 Online:2015-11-30 Published:2015-11-30

Abstract: Objective To construct the alpha-fetroprotein(AFP) mediated lentivirus in order to silence the expression of insulin like growth factor 1 receptor (IGF1R) and over-express p53 (Wtp53), and to explore the effect of lentivirus on the double target gene system (IGF1R and Wtp53)of liver cancer stem cells CD45-CD90+ and growth of liver cancer in vivo. Methods Liver cancer stem cells CD45-CD90+ were isolated from Hep3B cells. The lentivirus carrying the fusion genes of AFP-Wtp53-pPRIME-miR30-shRNA-IGF1R were used for in vitro transfection. Western blotting and RT-PCR were used to detect the expressions of wtp53 and IGF1R in the cells. The transplanted tumor model was established in BALB-C nude mice and then randomly divided into the blank control group (axillary inoculation with CD45-CD90+ cells), empty vector control group (axillary inoculation with CD45-CD90+ cells infected with empty vector pPRIME)and experimental group(axillary inoculation with CD45-CD90+ cells infected with AFP-Wtp53-pPRIME-miR30-shRNA-IGF1R). The IGF1R expression, microvessel density and apoptosis in each group were detected. Results The anti-AFP mediated lentivirus was successfully constructed. The Results of qRT-PCR and Western blotting showed that the recombinant lentivirus can over-express Wtp53 and interfere the expression of IGF1R. In vivo experiments showed that in comparison with the blank control group and empty vector control group, there were longer incubation period, smaller tumor volume, lower level of IGF1R and microvessel density in tumor tissue and higher apoptosis index in experimental group(P<0.05). Conclusion Anti-AFP mediated lentivirus targeted double target gene system of CD45-CD90+ cells presents high efficiency and specificity, playing a role in the growth of liver cancer stem cells and inhibits the growth of liver cancer cells.

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