本文選題：乳腺癌干細胞 + TF15基因��； 參考：《中國人民解放軍醫(yī)學院》2017年碩士論文

【摘要】：目的:乳腺癌是女性常見的惡性腫瘤,通過手術切除癌灶結合放化療或生物治療的方法約50%的患者可被治愈,但仍然有部分患者在治療5年內出現復發(fā)或轉移最終導致治療失敗,因此復發(fā)和轉移成為乳腺癌患者死亡的重要因素。研究已證實,乳腺癌細胞中存在小部分具有自我更新、多向分化潛能及高致瘤性的細胞亞群,即乳腺癌干細胞。乳腺癌干細胞可干預乳腺癌的發(fā)生、發(fā)展、侵襲等多個環(huán)節(jié)從而影響臨床療效。TF15基因為脊柱動物特異性同源異型基因,主要功能為一種編碼結構特異性蛋白質。TF15基因含有三個同源結構域,同源結構域可以與DNA結合,在動物體發(fā)育早期便起始作用,與胚胎干細胞的形成、神經系統發(fā)育以及多種先天性基因缺陷病有關,但TF15基因對乳腺癌的影響尚未明確。本研究通過分析TF15基因對乳腺癌干細胞生長、侵襲特性的影響,為臨床診治乳腺癌提供了新方向。方法:1、應用流式細胞儀計數、懸浮培養(yǎng)及裸鼠成瘤試驗檢測TF15基因對乳腺癌干細胞含量、生長特性的影響;2、通過RT-PCR、免疫熒光染色等試驗驗證TF15與上皮細胞向間質細胞轉化(EMT: epithelial mesenchymal transition)是否存在相關性;3、Western blot、RT-PCR等試驗篩選TF15影響細胞干性的轉錄因子。結果:1、過表達TF15后,通過流式細胞技術檢測到乳腺癌干細胞的含量明顯增多;懸浮培養(yǎng)結果顯示高表達TF15細胞形成微球體的能力增強,在小鼠體內TF15可增加乳腺癌致瘤能力;2、TF15過表達后EMT相關基因N-cadherin和Vinmintin轉錄及蛋白水平表達升高而E-cadherin降低,但TF15不影響此三個基因的啟動子活性,提示TF15過表達間接促進乳腺癌細胞上皮向間質轉化現象;3、通過RT-PCR及Western blot篩選出TF15相關轉錄因子Notch-1及C-myc。結論:在乳腺癌中,TF15基因可能通過Notch-1及C-myc增加乳腺癌干細胞含量并增強乳腺癌細胞致瘤能力。
[Abstract]:Objective: breast cancer is a common malignant tumor in women. About 50% of the patients can be cured by surgical resection combined with radiotherapy, chemotherapy or biotherapy. However, there are still some patients who have recurred or metastasized within 5 years after treatment, which lead to the failure of treatment, so recurrence and metastasis become an important factor in the death of breast cancer patients. It has been confirmed that there are a small number of breast cancer cells with self-renewal, multi-differentiation potential and highly tumorigenic subsets, namely breast cancer stem cells. Breast cancer stem cells can interfere with the occurrence, development and invasion of breast cancer, thus affecting clinical efficacy. TF15 gene is specific homologous gene of spine animal. The main function is that a structure-specific protein. TF15 gene contains three homologous domains, which can bind to DNA and begin to function with embryonic stem cells at the early stage of animal development. Nervous system development and many congenital genetic defects are related, but the effect of TF15 gene on breast cancer is not clear. By analyzing the effect of TF15 gene on the growth and invasion of breast cancer stem cells, this study provides a new direction for clinical diagnosis and treatment of breast cancer. Methods: the content of TF15 gene on breast cancer stem cells was detected by flow cytometry, suspension culture and tumorigenesis test in nude mice. The influence of growth characteristics was investigated by RT-PCR and immunofluorescence staining. Whether there was a correlation between TF15 and the transformation of epithelial cells to stromal cells (EMT: epithelial mesenchymal transition) was detected by RT-PCR. The transcription factors of TF15 affecting cell dryness were screened by RT-PCR. Results after overexpression of TF15, the amount of breast cancer stem cells was significantly increased by flow cytometry, and suspension culture showed that the ability of high expression TF15 cells to form microspheres was enhanced. TF15 increased the expression of N-cadherin and Vinmintin, but decreased E-cadherin, but TF15 did not affect the promoter activity of these three genes. These results suggest that TF15 overexpression can indirectly promote the epithelium to mesenchymal transformation of breast cancer cells. The transcription factors Notch-1 and C-myc were screened by RT-PCR and Western blot. Conclusion: tTF15 gene may increase the number of breast cancer stem cells and enhance the tumorigenicity of breast cancer cells through Notch-1 and C-myc in breast cancer.
【學位授予單位】：中國人民解放軍醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R737.9

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本文編號：1998531