TF15基因对乳腺癌干细胞生长特性的调控作用与机制探讨

发布时间：2018-06-09 02:58

本文选题：乳腺癌干细胞 + TF15基因　；参考：《中国人民解放军医学院》2017年硕士论文

【摘要】：目的:乳腺癌是女性常见的恶性肿瘤,通过手术切除癌灶结合放化疗或生物治疗的方法约50%的患者可被治愈,但仍然有部分患者在治疗5年内出现复发或转移最终导致治疗失败,因此复发和转移成为乳腺癌患者死亡的重要因素。研究已证实,乳腺癌细胞中存在小部分具有自我更新、多向分化潜能及高致瘤性的细胞亚群,即乳腺癌干细胞。乳腺癌干细胞可干预乳腺癌的发生、发展、侵袭等多个环节从而影响临床疗效。TF15基因为脊柱动物特异性同源异型基因,主要功能为一种编码结构特异性蛋白质。TF15基因含有三个同源结构域,同源结构域可以与DNA结合,在动物体发育早期便起始作用,与胚胎干细胞的形成、神经系统发育以及多种先天性基因缺陷病有关,但TF15基因对乳腺癌的影响尚未明确。本研究通过分析TF15基因对乳腺癌干细胞生长、侵袭特性的影响,为临床诊治乳腺癌提供了新方向。方法:1、应用流式细胞仪计数、悬浮培养及裸鼠成瘤试验检测TF15基因对乳腺癌干细胞含量、生长特性的影响;2、通过RT-PCR、免疫荧光染色等试验验证TF15与上皮细胞向间质细胞转化(EMT: epithelial mesenchymal transition)是否存在相关性;3、Western blot、RT-PCR等试验筛选TF15影响细胞干性的转录因子。结果:1、过表达TF15后,通过流式细胞技术检测到乳腺癌干细胞的含量明显增多;悬浮培养结果显示高表达TF15细胞形成微球体的能力增强,在小鼠体内TF15可增加乳腺癌致瘤能力;2、TF15过表达后EMT相关基因N-cadherin和Vinmintin转录及蛋白水平表达升高而E-cadherin降低,但TF15不影响此三个基因的启动子活性,提示TF15过表达间接促进乳腺癌细胞上皮向间质转化现象;3、通过RT-PCR及Western blot筛选出TF15相关转录因子Notch-1及C-myc。结论:在乳腺癌中,TF15基因可能通过Notch-1及C-myc增加乳腺癌干细胞含量并增强乳腺癌细胞致瘤能力。
[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.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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