携带11r-p53和mgm-csf基因载体的构建及其表达和对肿瘤细胞作用的分析-construction and expression of vector carrying 11r - p53 and mgm - csf genes and analysis of their effects on tumor cells.docxVIP

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2018-08-10 发布于上海
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携带11r-p53和mgm-csf基因载体的构建及其表达和对肿瘤细胞作用的分析-construction and expression of vector carrying 11r - p53 and mgm - csf genes and analysis of their effects on tumor cells.docx

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携带11r-p53和mgm-csf基因载体的构建及其表达和对肿瘤细胞作用的分析-construction and expression of vector carrying 11r - p53 and mgm - csf genes and analysis of their effects on tumor cells

PAGE PAGE IV 携带 11R-p53 和 mGM-CSF 基因载体的构建及其表达和对肿瘤细胞作用的研究英文摘要 The structure and expression of adenoviruses carrying 11R-P53 and mGM-CSF and it inbitionaleffect towards tumor tissue. Abstract Immunotherapy and genetherapy of the hepatocellular carcinoma has currently shown potential effect and good future. Antioncogene P53 and gene GM-CSF was recombined into the same adenovirus vector in this text. Before large-scale preparation of plasmid DNA, recombined adenovirus vector was transfect into packaging cell. The recombinant gene was renamed as SG655-mGMP. The potential anti-tumor effect was identified by testing the expression of desired gene in vitro and in vivo. SG655-mGMP was expect to show potent effect towards tumor cell which can kill or stop the growth of the tumor cell through gene level and cell immunology level. Aim: To construct an adenoviral vector carrying 11R-P53 and mGM-CSF gene and also detect its expression in tumor cell. Method. Recombination these genes to the area of E1B-55KD by combined bisulfite restr iction assay. Adenovirus packaging and prepare plasmid in quantity after DNA sequencing, and named as SG655-Mgmp. Virus titer was tested by TCID50, and transfection in vitro was conducted to 293 cells, Huh7 cells and Hepa1-6 cells using SG655-mGMP(MOI=5) and then teste the expression of 英文摘要携带 11R-p53 和 mGM-CSF 基因载体的构建及其表达和对肿瘤细胞作用的研究 the desired gene by western blot and ELESA in vivo. Forty C57BL/6 mouse was injected with Hepa1-6 during logarithmic phase, 5×106 cells per mouse. The mouse were distributed into four groups named A,B,C and D, and was injected with SG655-mGMP, SG7605-11R-P53, SG7605-mGM-CSF and normal saline (NS). The expression of the desired gene was testing by Immunohistochemistry. Result. The SG655-mGMP was structured successfully and the titer is 7.3×109PFU/ml. And the desired gene can express in vivo and in vitro. Keyword Biological therapy；Hepatocellular carcinoma；P53；GM-CSF Written by: Zhang Xiaokang Supervised by:Yang Jiahe