δ-三烯生育酚抗鼻咽癌的生物活性及其多靶点机制

In previous research, delta-tocotrienol has been proved to inhibit proliferation and induce apoptosis of nasopharyngeal carcinoma CNE2 and 5-8F cell lines, sharply down-regulate the expression of 3-hydroxy-3-methylglutaryl- coenzyme A (HMG-CoA) reductase and nuclear transcription factor(NF-κB) which play important role in tumor cell signaling pathways. To reveal the biological activity of delta-tocotrienol on nasopharyngeal carcinoma and its mechanism, this project will focus on three works: 1) Constructing HMG CoA reductase and/or NF k-B silencing nasopharyngeal carcinoma cell lines. 2) Exploring the effects of delta-tocotrienol on proliferation, apoptosis, angiogenesis, invasion and metastasis of HMG CoA reductase and/or NF k-B silencing nasopharyngeal carcinoma cell lines. 3) Analyzing the signal transduction pathways and signal molecules with corresponding gene chips to detect differential expression genes caused by delta-tocotrienol, checking these genes by real-time fluorescence quantitative PCR and Western hybridization. This project will not only make systematic understanding of biological activity of delta-tocotrienol on nasopharyngeal carcinoma, but also help to ascertain its working signaling pathways and molecular targets, and look out new medicine and targets for fighting nasopharyngeal carcinoma safely and effectively.

前期研究发现δ-三烯生育酚显著抑制鼻咽癌CNE2和5-8F细胞增殖，诱导其凋亡，进一步发现δ-三烯生育酚明显下调肿瘤细胞信号通路中起重要作用的羟甲基戊二酸单酰CoA (HMG CoA) 还原酶和核转录因子（NF-κB），为了探明其抗鼻咽癌的生物学作用与机制,本项目拟从三方面展开研究：1）构建HMG CoA还原酶和/或NF-κB沉默的鼻咽癌细胞株。2）研究δ-三烯生育酚对鼻咽癌细胞、HMG CoA还原酶和/或NF-κB沉默的鼻咽癌细胞增殖、凋亡、血管生成和侵袭转移的生物学作用。3）进一步采用信号转导通路基因芯片、实时荧光定量PCR和Western 杂交技术，筛选和确定δ-三烯生育酚抗鼻咽癌的信号转导通路及作用靶点。项目不仅有助于系统地阐明δ-三烯生育酚抗鼻咽癌的生物学活性，探明其抗鼻咽癌作用的信号通路和分子靶点，而且有望发现新的抗鼻咽癌药物和药物靶点，为安全有效地防治鼻咽癌提供实验依据。

本课题采用体外和体内实验研究δ-三烯生育酚抗鼻咽癌CNE2和5-8F细胞的生物学活性，并研究了δ-三烯生育酚对NF-κB和HMG CoA还原酶（HMGR）沉默的CNE2和5-8F细胞的生物学作用。实验发现δ-三烯生育酚对鼻咽癌细胞及NF-κB沉默的鼻咽癌细胞的增殖具有抑制作用，诱导其凋亡；δ-三烯生育酚抑制鼻咽癌细胞及NF-κB沉默的鼻咽癌细胞移植瘤生长，诱导移植瘤癌细胞凋亡，具有较强的抗鼻咽癌作用。基因表达谱芯片研究发现，δ-三烯生育酚可改变多个细胞增殖、凋亡和侵袭转移相关基因的表达，δ-三烯生育酚对NF-κB沉默后的鼻咽癌细胞的基因和信号分子表达仍有作用。荧光定量RT-PCR检测发现，δ-三烯生育酚对NF-κB和HMGR沉默的鼻咽癌细胞的E-钙粘附蛋白（E-cadherin）、羧基末端结合蛋白（CTBP）等其他基因的表达有影响。δ-三烯生育酚抗鼻咽癌的作用不仅可通过抑制NF-κB的活性，还可能通过影响E-cadherin和CTBP等其他基因的表达发挥作用。本课题研究获得了较完整的δ-三烯生育酚抗鼻咽癌的实验数据，研究成果将为天然药物抗鼻咽癌的研究和应用提供有意义的参考和思路。