SV40Tag转基因大鼠胰岛细胞瘤模型的建立、鉴定以及发病机理的初步研究

发布时间：2018-04-04 23:28

本文选题：SV40Tag　切入点：胰岛细胞瘤　出处：《扬州大学》2008年硕士论文

【摘要】： Simian Virus40(SV40)属于多瘤病毒属,是一种致瘤病毒,其早期基因编码产物大T抗原,是一种多功能调节蛋白,可以结合抑癌基因(如p53、pRb)与之相互作用,刺激DNA的复制,从而促进细胞的增值,引起正常细胞发生转化。利用这一特性,建立了大量的肿瘤转基因动物模型,包括胰腺癌、骨肉瘤、膀胱癌、肝癌等。而关于SV40Tag的转基因胰岛细胞瘤模型报道较少。胰岛细胞瘤也称胰腺内分泌肿瘤,临床上多出现低血糖症状,但是此症状并不特异,并且由于其位置隐匿,多在晚期发现,并已经激发产生多种病症,早期诊断,治疗十分困难,所以关于胰岛细胞瘤的相关研究也较少,目前其发病机理尚不完全清楚。本研究通过SV40Tag转基因大鼠的建立,以期得到可稳定遗传的胰腺瘤大鼠,为胰腺瘤的深入研究以及SV40Tag致瘤机理的研究提供大量的动物模型。利用pBC-SV40Tag重组质粒对SD大鼠进行雄原核显微注射,经PCR、RT-PCR和Western Blotting检测证实获得了阳性转基因大鼠,80%SV40Tag阳性转基因大鼠在6~9月龄出现胰腺肿瘤。通过解剖、组织病理学、免疫组织化学分析以及临床血糖测试,证明该肿瘤源自胰腺的胰岛细胞,与人的胰岛素细胞瘤极为相似,确定为胰岛细胞瘤。根据本实验室早期相关研究结果,利用real-time PCR以及免疫共沉淀结合Western Blotting对该肿瘤模型中IGF-IR信号通路上的重要蛋白IRS-1进行分析,发现其与SV40Tag存在相互作用,在SV40Tag表达的细胞中,异常增加,并且随SV40Tag进入细胞核中。综上所述,本研究成功建立了SV40Tag转基因大鼠,得到了类似于人的胰岛细胞瘤模型,并对其发病机理进行了初步探讨,这将对SV40Tag的致瘤机理的研究以及胰岛细胞瘤的预防、临床诊断以及治疗具有重要作用。
[Abstract]:Simian virus 40 (SV40) belongs to the genus polytumour virus. It is an early gene encoding a large T antigen and a multifunctional regulatory protein, which can interact with tumor suppressor genes (such as p53 pRb) and stimulate the replication of DNA.This promotes the cell increment, causes the normal cell to produce the transformation.Using this characteristic, a large number of tumor transgenic animal models have been established, including pancreatic cancer, osteosarcoma, bladder cancer, liver cancer and so on.However, there are few reports about SV40Tag's transgenic islet cell tumor model.Islet cell tumor is also called pancreatic endocrine tumor. It often appears hypoglycemia in clinic, but the symptom is not specific, and because its location is hidden, it is found in the late stage, and has already stimulated many kinds of diseases, early diagnosis,Treatment is very difficult, so there are few studies on islet cell tumor, and the pathogenesis of islet cell tumor is not completely clear.In this study, SV40Tag transgenic rats were established in order to obtain stable and hereditary pancreatic tumor rats, and to provide a large number of animal models for the further study of pancreatic tumor and the study of the mechanism of SV40Tag tumorigenesis.The male prokaryotic cells of SD rats were microinjected with pBC-SV40Tag recombinant plasmid. The pancreatic tumors were detected by PCR and Western Blotting in 80 SV40 tag positive transgenic rats at the age of 69 months.By dissection, histopathology, immunohistochemical analysis and clinical blood glucose test, it was proved that the tumor originated from pancreatic islet cells and was identified as islet cell tumor, which is very similar to human insulinoma.According to the results of early studies in our laboratory, real-time PCR and immunoprecipitation combined with Western Blotting were used to analyze the important protein IRS-1 in IGF-IR signaling pathway in this tumor model. It was found that IRS-1 interacted with SV40Tag in SV40Tag cells.Abnormal increase, and with SV40Tag into the nucleus.In conclusion, SV40Tag transgenic rats were successfully established, and a human islet cell tumor model was obtained, and its pathogenesis was preliminarily discussed, which will contribute to the study of the tumorigenic mechanism of SV40Tag and the prevention of islet cell tumor.Clinical diagnosis and treatment play an important role.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R736.7;R-332

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