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The Effects of Lentivirus Mediated bcr/abl RNAi on Chronic Granulocytic Leukemia Proliferation

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Tutor: HaoXinBao;CaoXianYing
School: Hainan University
Course: Biochemical Engineering
Keywords: lentivirus,bcr/abl gene,P210bcr/abl,cell proliferation
CLC: R733.72
Type: Master's thesis
Year:  2014
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Objective:To study the effects of lentivirus mediated bcr/abl RNAi on the viability of K562cell line and prepare for the further study and clinic treatment.Methods:We transfected the lentivirus vectors containing bcr/abl RNAi to K562cell line, and selected stable transfectants. To assess the RNAi efficiency, we used Real-time PCR to test the transcription of bcr/abl, while western blotting to test the expression of P210, which is coded by bcr/abl. Cell viability was detected by CCK-8assay, clonogenic assay, BrdU incorporation assay and cell cycle analysis. Finally test the in vivo tumorigenesis in node mice.According to the results of RT-PCR and western blot, transcription of bcr/abl gene and expression of P210both decreased in the transfected cells. As the CCK-8assay shows, proliferation of bcr/abl RNAi transfected cells was significantly inhibited compared with negative control. Clonogenic assay showed the colony number of the experiment group were only about40%of the control group. In conclusion, inhibition of bcr/abl gene expression leads to the decrease of the cell proliferation about60%. Cell cycle analysis suggested that the cells in G0/G1phase are much more in experimental group than control group. And also, BrdU incorporation level is much lower in the experimental group than control group. And also, in vivo study shows that experiment group has lower tumorigenic ability compared with control group.Conclusion:Bcr/abl RNAi mediated by lentivirus can strikingly inhibit the proliferation of K562cells, which makes the bcr/abl RNAi a promising way to cure leukemia.
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