本文選題：鼻咽癌 + 雞胚絨毛尿囊膜��； 參考：《廣西醫(yī)科大學(xué)》2010年博士論文

【摘要】： 鼻咽癌是我國高發(fā)的惡性腫瘤,而廣西是全國鼻咽癌的第二高發(fā)區(qū)。在過去二十年中,廣西的鼻咽癌發(fā)病率及死亡率幾乎無明顯改善。建立鼻咽癌體內(nèi)動物實(shí)驗(yàn)?zāi)Ｐ褪茄芯勘茄拾┑闹匾侄?也是我們急需解決的課題之一。目前常用的鼻咽癌動物模型是裸鼠移植瘤模型,其實(shí)驗(yàn)條件相對較高,且實(shí)驗(yàn)周期長,費(fèi)用昂貴。而雞胚絨毛尿囊膜(CAM)模型以簡單、方便、價廉、快速等優(yōu)點(diǎn)已日益成為腫瘤研究工作者常用的一個活體模型。我們的目的是建立基于雞胚絨毛尿囊膜的高效、實(shí)用、經(jīng)濟(jì)的鼻咽癌體內(nèi)動物實(shí)驗(yàn)?zāi)Ｐ?為將來進(jìn)一步研究鼻咽癌細(xì)胞的浸潤轉(zhuǎn)移機(jī)制、抗腫瘤藥物篩選、臨床前期評價乃至腫瘤個體治療方案等建立技術(shù)平臺。 經(jīng)過反復(fù)摸索,我們對建立該移植瘤模型的方法和條件進(jìn)行了優(yōu)化：根據(jù)雞胚孵化特點(diǎn),確定實(shí)驗(yàn)的開始時間為雞胚孵至第8日齡時,最遲的終止時間為雞胚孵至第14日齡；確定以“開窗法”為我們獲取CAM的方法；以硅膠圈為我們接種腫瘤細(xì)胞的載體；完善并熟練掌握了雞胚“開窗”的技術(shù),使雞胚“開窗”成功率接近100%；確定鼻咽癌細(xì)胞的最佳接種細(xì)胞數(shù)。通過以上條件的優(yōu)化,我們成功建立和完善了鼻咽癌雞胚CAM移植瘤模型。 同時,我們發(fā)現(xiàn),鼻咽癌細(xì)胞在CAM移植瘤形成的過程中可誘導(dǎo)宿主新生血管形成,形成的新生血管以移植瘤為中心呈放射狀的排列。我們探索了通過測算尿囊膜／血管面積比來定量研究新生血管的方法,并發(fā)現(xiàn),隨著接種細(xì)胞數(shù)的增多,新生血管也增多。我們構(gòu)建了標(biāo)記GFP的鼻咽癌細(xì)胞株,接種于CAM,通過激光共聚焦顯微鏡觀察GFP標(biāo)記的鼻咽癌細(xì)胞的侵襲行為。通過此模型我們可觀察到癌細(xì)胞浸潤并突破基底膜及血道轉(zhuǎn)移的早期過程。我們還探討了CAM模型用于研究鼻咽癌細(xì)胞遠(yuǎn)處轉(zhuǎn)移的可行性。通過絕對定量real-time熒光定量PCR檢測人β-globin基因,我們在接種鼻咽癌細(xì)胞5天的雞胚心肺組織內(nèi)檢測到了微量的人癌細(xì)胞。在此基礎(chǔ)上,我們驗(yàn)證了兩株不同轉(zhuǎn)移潛能鼻咽癌細(xì)胞株的遠(yuǎn)處轉(zhuǎn)移能力差異,并證實(shí)CAM模型可以用于腫瘤轉(zhuǎn)移能力的預(yù)測。 總之,本研究首次建立了鼻咽癌雞胚絨毛尿囊膜移植瘤模型,并從形態(tài)學(xué)、分子生物學(xué)等方面闡述該活體模型應(yīng)用于鼻咽癌新生血管的定量研究、鼻咽癌早期浸潤侵襲的生物學(xué)行為的觀察以及鼻咽癌遠(yuǎn)處轉(zhuǎn)移潛能的評價。本課題工作為進(jìn)一步開展鼻咽癌的腫瘤生物學(xué)相關(guān)研究奠定了良好的工作基礎(chǔ),提供了良好的技術(shù)平臺。
[Abstract]:Nasopharyngeal carcinoma is a high incidence of malignant tumor in China, and Guangxi is the second high incidence area of nasopharyngeal carcinoma in China. In the past twenty years, the incidence and mortality of nasopharyngeal carcinoma in Guangxi have almost no obvious improvement. The animal model of nasopharyngeal carcinoma (nasopharyngeal carcinoma) is a model of nude mice. The experimental conditions are relatively high, and the experimental period is long and the cost is expensive. The chicken embryo chorioallantoic membrane (CAM) model has become a living model commonly used by the cancer researchers for the advantages of simple, convenient, cheap and fast. The animal experimental model of efficient, practical and economical nasopharyngeal carcinoma in vivo is a technical platform for further study of the mechanism of invasion and metastasis of nasopharyngeal carcinoma cells, screening of antitumor drugs, pre clinical evaluation and even individual treatment of tumor.
After repeated exploration, we have optimized the method and conditions for establishing the model of the transplanted tumor. According to the characteristics of chicken embryo hatching, it is determined that the start time of the experiment is eighth days of age when the chicken embryo is hatched to the age of eighth days, the last time for the chicken embryo is hatched to fourteenth days of age. The carrier of the tumor cells was planted, and the technique of "opening the window" of the chicken embryo was perfected and mastered to make the success rate of the chicken embryo "open window" close to 100%, and to determine the optimum number of inoculation cells for nasopharyngeal carcinoma cells. Through the optimization of the above conditions, we successfully established and perfected the CAM model of the chicken embryo of nasopharyngeal carcinoma.
At the same time, we found that nasopharyngeal carcinoma cells can induce the formation of the host neovascularization in the CAM transplant tumor, and the new blood vessels form a radial arrangement in the center of the transplanted tumor. We explored the method of quantitative study of the neovascularization by measuring the allantoic membrane / vascular area ratio, and found that as the number of inoculated cells increases, The neovascularization was also increased. We constructed a nasopharyngeal carcinoma cell line marked GFP, inoculated in CAM, and observed the invasion behavior of GFP labeled nasopharyngeal carcinoma cells by laser confocal microscopy. Through this model, we can observe the infiltration of the cancer cells and break through the early process of the basement membrane and the metastasis of the blood channel. We also explored the CAM model for the study of the nose. The feasibility of distant metastasis of the pharyngeal carcinoma cells was detected by the absolute quantitative real-time fluorescence quantitative PCR detection of human beta -globin gene. We detected a small number of human cancer cells in the chicken embryo cardiopulmonary tissues of 5 days of nasopharyngeal carcinoma cells. On this basis, we verified the difference in the distant metastasis ability of two different metastatic potential nasopharyngeal carcinoma cells. It is confirmed that the CAM model can be used to predict tumor metastasis.
In conclusion, this study was the first time to establish a model of nasopharyngeal choriochorioallantoic membrane transplantation, and the quantitative study on the application of the living model to nasopharyngeal cancer neovascularization, the biological behavior of nasopharyngeal invasion and the evaluation of the distant metastasis potential of nasopharyngeal carcinoma from morphology, molecular biology and other aspects. Further research on tumor biology in nasopharyngeal carcinoma has laid a good foundation for work and provided a good technical platform.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2010
【分類號】：R739.63

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 鄭秀海,王曙光,倪嘉,李昆,韓本立;人膽管癌雞胚移植瘤模型的建立及其生物學(xué)特性的研究[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2003年09期

2 冷雷;劉騰飛;黃仲曦;謝衛(wèi)兵;姚開泰;;鼻咽癌淋巴結(jié)轉(zhuǎn)移動物模型的建立及其轉(zhuǎn)移相關(guān)標(biāo)簽基因的篩選[J];南方醫(yī)科大學(xué)學(xué)報(bào);2008年09期

3 吳敬波;文慶蓮;范娟;郎錦義;王艷萍;劉學(xué)旭;;拓?fù)涮婵祵Ρ茄拾┓派湓雒舻捏w內(nèi)實(shí)驗(yàn)[J];中國耳鼻咽喉頭頸外科;2007年02期

4 周蓉蓉;陳嘉;肖志強(qiáng);;E1A基因?qū)θ吮茄拾﹦游锬Ｐ头派湓雒舻膶?shí)驗(yàn)研究[J];中南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年08期

5 李曉華;詹志榮;孫建設(shè);曾斌;;鼻咽癌細(xì)胞株裸鼠肝異位種植瘤肺轉(zhuǎn)移動物模型的建立[J];江西醫(yī)學(xué)院學(xué)報(bào);2008年02期

6 楊麗華,王凱峰,周軼平,李瑪琳;雞胚尿囊膜模型血管生成定量指標(biāo)的研究[J];昆明醫(yī)學(xué)院學(xué)報(bào);2004年02期

7 陳昕;文慶蓮;鄧紅梅;劉安敏;;裸鼠不同部位皮下接種鼻咽癌瘤塊成瘤特點(diǎn)比較[J];瀘州醫(yī)學(xué)院學(xué)報(bào);2006年03期

8 阿斯楞;仝林虎;蘇秀蘭;;腫瘤血管生成的分子機(jī)制[J];內(nèi)蒙古醫(yī)學(xué)雜志;2006年03期

9 吳四海;俞晨杰;;鼻咽癌移植瘤動物模型的建立及其腫瘤生物學(xué)特性觀察[J];齊齊哈爾醫(yī)學(xué)院學(xué)報(bào);2007年18期

10 肖志強(qiáng)，，姚開泰，劉薇，盧光　;攜帶EB病毒潛伏膜蛋白基因轉(zhuǎn)基因小鼠的制備[J];生物化學(xué)與生物物理進(jìn)展;1995年02期

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