慢病毒介导BMPRⅡ基因沉默对人肝癌移植瘤生长的影响-肿瘤学专业毕业论文.docxVIP

Abst Abstract IV IV ABSTRACT Objective： To study the effects of the recombinant lentiviral vector for RNA interference (RNAi) of BMPR II gene on the expression of BMPR II gene and the growth of subcutaneous tumor of hepatocellular carcinoma in nude mice. Methods： Construct one shRNA-BMPR II plasmid and a negative control plasmid, The mRNA and protein levels of BMPR II in group of HepG2-control、HepG2-shRNA(- ) 、HepG2- shRNA(+) were determined by real-time fluorescence quantitative PCR and western blotting and make sure the plasmid of HepG2- shRNA(+) obvious interference effect for lentivirus packaging. BALB/C nude mice were inoculated subcutaneously with HepG2 cells to establish the subcutaneous tumor model of human hepatocellular carcinoma. The mice were randomly divided into experimental group, negative control group and blank control group. BMPR II shRNA lentiviral particles, lentiviral particles which carrying no effective sequence or NS(0.2 ml per time) were injected individually every two days. The volume of tumor of mice were measured. and the expression of BMPR II was detected by Western bolt and immumohistochemical （IHC）. Results： The expression level of BMPR II mRNA and protein was detected by qRT PCR and Western bolt show the BMPR II shRNA(+) group were obviously decreased(P0.05). The mean rate of tumor growth in the experimental group was much slower than in the negative control group and blank control group(P0.05). The dissected tumors in the blank control group and the negative control group had volumes of （1255.2±130.05） mm3 and （1274.6±133.02）mm3, whereas in the experimental group, the tumors had a volume of （328.6±87.72） mm3.And BMPR PAGE PAGE VI II protein was detected by Western bolt and immumohistochemical show the experimental group were obvious decreased(P0.05). Conclusion： Intra-tumor injection of BMPR II miRNA lentiviral vector has a significantly inhibitory effect on the growth of subcutaneous tumor of hepatocel