TIM-3 促上皮-间质转化诱导骨肉瘤侵袭及转移的分子机制研究

项目名称： TIM-3 促上皮-间质转化诱导骨肉瘤侵袭及转移的分子机制研究

项目编号： No.61461004

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 无线电电子学、电信技术

项目作者： 尚永军

作者单位： 赤峰学院

项目金额： 47万元

中文摘要： 前期研究我们发现免疫分子Tim-3特异表达于人骨肉瘤组织并与上皮-间质转化(EMT)标志物共表达，提示Tim-3可能通过诱导EMT促骨肉瘤的侵袭及转移（Oncol Lett. 2013:490-494）。然而Tim-3促进EMT的分子机制尚未明确。本研究拟收集大样本量的骨肉瘤临床标本，分析Tim-3表达与骨肉瘤术后预后的相关性；在人骨肉瘤MG-63细胞表达Tim-3，采用qPCR、EMSA及Western-blot等技术分析Tim-3信号下调ESRP/FGFL2b导致EMT进程的分子机制；并应用划伤愈合及Transwell培养技术分析TIM-3对细胞侵袭能力的影响；最后建立人MG-63细胞的SCID小鼠移植瘤模型，对比肿瘤大小、小鼠存活及EMT进程等指标，证实TIM-3可通过EMT作用增强癌细胞转移。本研究将阐明Tim-3促骨肉瘤转移的分子机制，对骨肉瘤的早期诊断及临床治疗有重要意义。

中文关键词： TIM-3；上皮间质转化；骨肉瘤；转移；细胞信号处理

英文摘要： Osteosarcoma （OS） is one of the serious carcinoma characterized by malignant invasion and metastasis that harm to human health. Nevertheless,the mechanism that induces tumor cell invasion and metastasis is unclear. Our previous work has found that the novel immunosuppressive molecule, T cell immunoglobulin and mucin-domain-containing molecule-3 (TIM-3),was specific expression in human osteosarcoma tissue. Interestingly, it co-expressed with some epithelial-mesenchymal transition (EMT) markers like Ecad, Smad, Snail et al., suggesting that TIM-3 should promote the pathogenesis of osteosarcoma via inducing the EMT process (Oncol Lett. 2013;6:490-494). In this project, the expression of TIM-3 in the osteosarcoma cell line-MG-63 cells should be overexpressed by DNA transfected technique, and, the capacity of tumor cell invasion should be analyzed by transwell experiments ex vivo. Moreover, TIM-3-MG63 and the control cells should be transferred into the SCID mouse to establish an xenograft tumor model, the tumor size, mouse survival, the EMT process and other parameters should be compared and analzyed, these tudies should further confirm that TIM-3 can enhance tumor metastasis through EMT in vivo. Finally, a cohort of clinical samples of OS shoule be cellected from our department and the expression of Tim-3 should be detected by immunohistochemistry, therefore, the relationships between Tim-3 expression and patients survival rate (survival times) should be analyzed.To measure how Tim-3 signal control EMT progress, overexpression of Tim-3 in MG-63 cells and the expression of EMT related molecules, including ESRP, FGFR2, N-cad, Slug et al should be analyzed by western-blot, qPCR and EMSA. In a word, this research will elucidate the molecular mechanism of Tim-3 promoting metastasis of laryngeal cancer, which might help us develop some novel methods for early diagnosis and treatment of OS.

英文关键词： TIM-3;EMT;carcinoma;transformation;cellsignalprocessing