本文選題：SV40Tag　切入點：胰島細胞瘤　出處：《揚州大學》2008年碩士論文

【摘要】： Simian Virus40(SV40)屬于多瘤病毒屬,是一種致瘤病毒,其早期基因編碼產(chǎn)物大T抗原,是一種多功能調(diào)節(jié)蛋白,可以結(jié)合抑癌基因(如p53、pRb)與之相互作用,刺激DNA的復(fù)制,從而促進細胞的增值,引起正常細胞發(fā)生轉(zhuǎn)化。利用這一特性,建立了大量的腫瘤轉(zhuǎn)基因動物模型,包括胰腺癌、骨肉瘤、膀胱癌、肝癌等。而關(guān)于SV40Tag的轉(zhuǎn)基因胰島細胞瘤模型報道較少。胰島細胞瘤也稱胰腺內(nèi)分泌腫瘤,臨床上多出現(xiàn)低血糖癥狀,但是此癥狀并不特異,并且由于其位置隱匿,多在晚期發(fā)現(xiàn),并已經(jīng)激發(fā)產(chǎn)生多種病癥,早期診斷,治療十分困難,所以關(guān)于胰島細胞瘤的相關(guān)研究也較少,目前其發(fā)病機理尚不完全清楚。本研究通過SV40Tag轉(zhuǎn)基因大鼠的建立,以期得到可穩(wěn)定遺傳的胰腺瘤大鼠,為胰腺瘤的深入研究以及SV40Tag致瘤機理的研究提供大量的動物模型。 利用pBC-SV40Tag重組質(zhì)粒對SD大鼠進行雄原核顯微注射,經(jīng)PCR、RT-PCR和Western Blotting檢測證實獲得了陽性轉(zhuǎn)基因大鼠,80%SV40Tag陽性轉(zhuǎn)基因大鼠在6~9月齡出現(xiàn)胰腺腫瘤。通過解剖、組織病理學、免疫組織化學分析以及臨床血糖測試,證明該腫瘤源自胰腺的胰島細胞,與人的胰島素細胞瘤極為相似,確定為胰島細胞瘤。 根據(jù)本實驗室早期相關(guān)研究結(jié)果,利用real-time PCR以及免疫共沉淀結(jié)合Western Blotting對該腫瘤模型中IGF-IR信號通路上的重要蛋白IRS-1進行分析,發(fā)現(xiàn)其與SV40Tag存在相互作用,在SV40Tag表達的細胞中,異常增加,并且隨SV40Tag進入細胞核中。 綜上所述,本研究成功建立了SV40Tag轉(zhuǎn)基因大鼠,得到了類似于人的胰島細胞瘤模型,并對其發(fā)病機理進行了初步探討,這將對SV40Tag的致瘤機理的研究以及胰島細胞瘤的預(yù)防、臨床診斷以及治療具有重要作用。
[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.
【學位授予單位】：揚州大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R736.7;R-332

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相關(guān)期刊論文 前1條

1 金凡,周淑貞,陶蓉芳,方茹蓉,項永兵,孫璐,高玉堂;上海市區(qū)惡性腫瘤發(fā)病趨勢1972～1994年[J];腫瘤;1999年05期

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本文編號：1712197