MTDH介导上皮-间质转化及肿瘤干细胞双表型调控头颈鳞癌转移的实验研究

MTDH, a novel oncogene, is associaed with cancer metastasis. The relationship between cancer stem cells and epithelial mesenchymal transition(EMT) has always been a hot spot in the process of cancer metastasis. We have previously found that MTDH expression was significantly associted with EMT, invasion and prognostic of squamous cell carcinoma of the head and neck(SCCHN). Also, MTDH could affect the expression of cancer stem cells related protein ABCG2. So we hypothesised that MTDH mediate SCCHN metastasis via inducing EMT and cancer stem cells double phenotype. In the present study, lentiviral vector plasmid will be used to mediate the expression of MTDH, and MTDH mediated metastasis of SCCHN will also be examed. Cancer stem-like cells (CSCs) will be isolated by flow cytometry and cell sorting, then its relationship between EMT will be measured. What's more, gene microarray will be employed to explore the relative signal pathway which participate in MTDH mediated SCCHN metastasis. This study will investigate the association between CSCs and EMT during the process of MTDH mediated SCCHN metastasis, and the key signal pathway which contribute to MTDH mediated SCCHN metastasis. The accomplishment of this study will elucidate the mechanism of MTDH mediated metastasis of SCCHN, which will be valuable for future study of SCCHN metastasis and molecular target therapy.

MTDH是新近发现的癌基因，与肿瘤侵袭转移密切相关。肿瘤转移过程中，肿瘤干细胞与上皮-间质转化(EMT)的关系一直是人们关注的热点。我们前期研究发现：MTDH表达与头颈鳞癌EMT、侵袭及预后密切相关，并影响肿瘤干细胞重要蛋白ABCG2表达。据此我们提出科学假设：MTDH通过介导EMT与肿瘤干细胞双表型调控头颈鳞癌侵袭转移。本项目通过慢病毒载体介导的质粒调控MTDH表达，检测其对头颈鳞癌侵袭转移的影响；流式细胞分选技术分选肿瘤干细胞样细胞（CSCs）并检测其与EMT的相互关系；利用基因芯片技术筛选鉴定MTDH调控头颈鳞癌侵袭转移的关键信号通路。本项目的成功实施将进一步阐明MTDH对头颈鳞癌侵袭转移的调控作用，为头颈鳞癌转移的分子机制研究及分子靶向治疗研究奠定新的理论基础。

MTDH是新近发现的癌基因，与肿瘤侵袭转移密切相关。肿瘤转移过程中，肿瘤干细胞与上皮-间质转化(EMT)的关系一直是人们关注的热点。我们前期研究发现：MTDH表达与头颈鳞癌EMT、侵袭及预后密切相关，并影响肿瘤干细胞重要蛋白ABCG2表达。据此我们提出科学假设：MTDH通过介导EMT与肿瘤干细胞双表型调控头颈鳞癌侵袭转移。. 本项目主要研究计划是将已构建的慢病毒介导的MTDH表达质粒（plenti-MTDH）及其对照质粒和shRNA质粒（plenti-MTDH-shRNA）及其对照质粒分别转染MTDH表达不同的头颈鳞癌细胞株，筛选出稳定高表达及沉默MTDH的细胞株；体内、体外实验验证MTDH对头颈鳞癌侵袭转移的调控；筛选出相关信号通路，进一步确定MTDH调控头颈鳞癌侵袭转移的关健信号通路；检测MTDH对头颈鳞癌CSCs表型的调控；验证MTDH介导的头颈鳞癌EMT及肿瘤细胞干细胞样细胞双表型的相互关系；检测MTDH在头颈鳞癌组织中的表达以及与临床资料的相关性。. 经过课题组成员的共同努力，证实MTDH在头颈鳞癌组织中高表达，并与肿瘤转移呈正相关，MTDH高表达组患者总生存率和无瘤生存率显著低于低表达组患者，可以作为判断预后的独立指标；建立了稳定高表达及沉默MTDH的细胞株，证实MTDH可以通过AKT信号通路调控头颈鳞癌的EMT、侵袭以及VEGF的表达。目前MTDH对头颈鳞癌CSCs表型的调控实验正在进行中。. 在该项目的资助下，共培养博士研究生8名，硕士研究生8名，其中毕业8名；共发表SCI论文6篇，中文核心期刊论文7篇；获湖南省科学技术进步奖二等奖1项。