CRAd.pEgrl-TRAIL-Smac联合放射对MDA-MB-231细胞杀伤效应的研究

发布时间：2018-03-06 14:10

本文选题：X射线　切入点：TRAIL　出处：《吉林大学》2013年博士论文　论文类型：学位论文

【摘要】：恶性肿瘤作为一种全身性疾病，其产生是由多基因、多途径及多因素参与逐步发展的复杂过程，单一疗法往往难以取得满意疗效，因此，合理地选择综合治疗的模式为有效地杀伤肿瘤细胞，彻底治愈癌症带来了希望。肿瘤基因-放射治疗的提出弥补了放射治疗与基因治疗各自的弊端，将二者有机地结合起来，发挥协同抗肿瘤作用，，既增强了肿瘤组织对放射治疗的敏感性、提高外源基因靶向转移和杀伤肿瘤细胞的效率，同时也可以降低照射剂量，缓解射线对肿瘤周围正常组织的损伤。本研究利用对肿瘤细胞具有靶向性的溶瘤腺病毒作为载体，利用Egr-1启动子可在辐射诱导下增强其下游目的基因表达的特性，以及TRAIL和Smac基因的协同促肿瘤细胞凋亡的作用，构建了携带TRAIL和Smac双基因的具有双重靶向的pAd.Egr1-TRAIL-Smac-hTERT-E1A-E1B-E1B55K的溶瘤腺病毒质粒，并在HEK293细胞中成功地将其包装成携带Egr-1启动子、TRAIL和Smac双基因的双重靶向溶瘤腺病毒CRAd.pEgr1-TRAIL-Smac，探讨在其联合X射线照射对MDA-MB-231细胞增殖及凋亡的影响，以及MDA-MB-231细胞凋亡途径中相关基因的mRNA和蛋白表达规律。实验结果表明，CRAd.pEgr1-TRAIL-Smac联合X射线照射对MDA-MB-231细胞具有明显的增殖抑制及其促凋亡作用，可使细胞中TRAIL、DR5、Smac、Cytc、caspase-8、-9和-3mRNA及蛋白表达增高。结果提示，CRAd.pEgr1-TRAIL-Smac联合放疗抑制MDA-MB-231细胞增殖和促进其凋亡的机制，涉及细胞S期和G2期阻滞及死亡受体途径与线粒体途径中TRAIL、DR5、Smac、Cytc、caspase-8、-9和-3等基因的相互作用。本研究结果为临床治疗肿瘤提供了重要的理论依据及实验基础。
[Abstract]:As a systemic disease, malignant tumor is produced by multiple genes, multiple pathways and many factors involved in the complex process of progressive development, monotherapy is often difficult to achieve satisfactory results, therefore, The reasonable choice of comprehensive therapy mode brings hope to kill tumor cells effectively and cure cancer completely. The proposal of tumor gene-radiotherapy makes up for the disadvantages of radiotherapy and gene therapy, and organically combines the two. The synergistic antitumor effect not only enhances the sensitivity of tumor tissue to radiotherapy, but also increases the efficiency of foreign gene targeting metastasis and killing tumor cells, and at the same time decreases the irradiation dose. Radiation damage to normal tissue around the tumor is alleviated. In this study, adenovirus targeting tumor cells was used as a vector to enhance the expression of downstream target genes by radiation induced by Egr-1 promoter, as well as the synergistic effect of TRAIL and Smac genes on the apoptosis of tumor cells. The adenovirus plasmids carrying TRAIL and Smac double genes with double targeting pAd.Egr1-TRAIL-Smac-hTERT-E1A-E1B-E1B55K were constructed. HEK293 cells were successfully packaged as double targeting adenovirus CRAd.pEgr1-TRAIL-Smaccontaining Egr-1 promoter trail and Smac. The effects of combined X-ray irradiation on the proliferation and apoptosis of MDA-MB-231 cells were investigated. The results showed that CRAd.pEgr1-TRAIL-Smac combined with X-ray irradiation could significantly inhibit the proliferation and promote apoptosis of MDA-MB-231 cells. The results suggested that CRAd.pEgr1-TRAIL-Smac combined with radiotherapy could inhibit the proliferation and promote the apoptosis of MDA-MB-231 cells. This study provides an important theoretical basis and experimental basis for clinical treatment of tumors, involving the interaction of cell S and G2 arrest and death receptor pathways with genes such as TRAILDR5, SmacCaspase-8, caspase-8, and 3 in the mitochondrial pathway.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R730.55

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