tlr3的激活抑制乳腺癌细胞mda-mb-231的增殖

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1、硕士学位论文学校代码：10023学号：$2009412TLR3的激活抑制乳腺癌细胞MDA．MB．231的增殖所院：姓名：指导教师：导师小组：学科专业：研究方向：完成日期：血液学研究所血液病医院徐淑霞韩忠朝教授中法实验室内科学．干细胞与肿瘤方向2012年5月目录第—部分TLR3的激活抑制乳腺癌细胞MDA-MB-231的增殖中文摘要·····················································1英文摘要·····································

2、················3前言························································-5材料与方法···················································7结果一·····················一··················一····一·····12讨论························································17参考文献······

3、·············一·······························20第二部分双链RNA引起大鼠胰岛素瘤细胞INS-I凋亡的相关研究摘要···一一一···一·”一一······一一一·一一·一····一·······一23前言························································24材料与方法··················································24结果一········一········

4、··-·································25讨论························································28参考文献·······························‘····················30综述一间充质干细胞与肿瘤相互作用分子机制的研究进展·······3l综述二病毒感染与糖尿病···································47附录一主要实验仪器和试剂··········

5、························56附录二主要溶液配制········································58附录三英文缩略语表·······一·······························60附录四在学期间发表文章····································6l致谢························································62北京协和医学院中国医学科学院硕士学位论文第一部分

6、TLR3的激活抑制乳腺癌细胞MDA．MB-231的增殖摘要研究背景及目的：Toll样受体(toll—likereceptors，TLRs)是广泛分布在免疫细胞尤其非特异免疫细胞以及某些体细胞甚至一些肿瘤细胞表面以及细胞内部的一类模式识别受体(patternrecognitionreceptor，PRR)，它们可以直接识别结合某些病原体或其产物所共有的高度保守的特定分子模式(pathogenassociatedmolecularpattern，PAMP)结构，及病原相关分子模式。人的TLRs一共有11个亚型，不同的亚

7、型识别不同的病原分子结构，TLR3主要识别病毒在复制过程中产生的双链RNA。TLR3在肿瘤的发生发展和肿瘤免疫中起到重要的作用，有报道称TLR3的激活能够促进某些肿瘤细胞的凋亡。本研究旨在探讨TLR3的特异性激动剂Poly(I：C)对恶性程度较强的乳腺癌纽胞MDA-MB-231增殖与凋亡的影响，并初步探讨其发挥作用的机制。方法：(1)反转录PCR检测Toll样受体各个亚型在mRNA水平上的表达；(2)CCK一8细胞活力试剂盒检测不同浓度Poly(I：C)对MDA-MB-231细胞增殖的影响；(3)流式细胞术用Ann

8、exinV—FITC／PI染色检测Poly(I：C)对MDA-MB一231细胞凋亡作用的影响：(4)实时定量反转录PCR检测相关细胞因子的表达变化；(5)WesternBlot检测Poly(I：C)引起细胞内信号通路蛋白的变化。1北京协和医学院中国医学科学院硕士学位论文结果：(1)在mRNA水平上，MDA-MB-231细胞TLRs各亚型，TLR8表达量比较低