【作者】 罗丹枫；

【导师】 马丁；

【作者基本信息】 华中科技大学 ， 妇产科学， 2009， 博士

【摘要】 目的：寻找、验证肿瘤靶向多肽TMTP1的受体，并对TMTP1靶向肿瘤作用的分子机理做初步探讨，寻找肿瘤治疗可能的新靶点。方法：利用Pulldown技术寻找TMTP1可能的受体，通过逆向亲和实验、生物信息学分析、竞争拮抗实验以及Realtime PCR和Western等方法从受体与配体的结合、受体的分布以及功能效应等方面加以验证；以MTT、FCM、Western-blot等方法验证TMTP1具有诱导凋亡以抑制肿瘤细胞生长及增殖的生物作用，并探讨TMTP1与受体结合后，发挥TMTP1生物效应的分子机理。结果：XPNPEP2和Hsp27是利用Pulldown技术寻找到的TMTP1可能的受体。其中XPNPEP2存在于细胞膜表面，可以和TMTP1稳固结合，并可以显著影响TMTP1与肿瘤细胞亲和和诱导凋亡的能力，其细胞、组织的表达分布也与TMTP1的筛选原理想吻合；Hsp27在细胞内与TMTP1结合，通过PI3K／Hsp27信号通路拮抗TMTP1诱导肿瘤细胞凋亡的生物学作用。结论：XPNPEP2是TMTP1的细胞膜受体，并介导了TMTP1进入肿瘤细胞内部；Hsp27是TMTP1的胞内结合蛋白，在拮抗TMTP1诱导肿瘤细胞凋亡过程中起着关键作用。更多还原

【Abstract】 Objective:To capture and verify the receptor of a tumor homing peptide TMTP1 is helpfulto know about the mechanism of tumor homing effect.The receptor may be a potentialtherapeutic target.Methods:The motheds of pull-down (affinity chromatography)method were used toseparate the membrane protein molecules interacting with TMTP1 and indetify the somekind(s)of protein(s)using the Spectral analysis from the mixture of proteins.The validationof receptor including following aspect:the subcellular localization,the distribution,thebinding and the functional testing.We also used MTT assay,FCM,western-blot to verifythe inhibiton on tumor cell proliferation induced by TMTP1 and to investigate themechanism.Results:XPNPEP2 and Hsp27 are candidate receptor captured by pull-down.XPNPEP2expresses in the cellular surface,it can bind TMTP1 strongly and shows a good results inthe functional test.Its distribution also coincides with the rules which we screens theTMTP1 by phage display.Hsp27 is a intracellular binding protein of TMTP1,it negativelyregulate the apoptosis induced by TMTP1 through the PI3K/Hsp27 pathway.Conclusions:XPNPEP2 is a cellular surface receptor of TMTP1 and mediate theendocytosis of TMTP1.Hsp27 is a intracellular binding protein of TMTP 1,it plays a keyrole in the apoptosis induced by TMTP 1.更多还原