Smac多肽、类Smac小分子对卵巢癌细胞的耐药性调节及其诱导凋亡的机制研究

项目名称： Smac多肽、类Smac小分子对卵巢癌细胞的耐药性调节及其诱导凋亡的机制研究

项目编号： No.30872739

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 刘培淑

作者单位： 山东大学

项目金额： 30万元

中文摘要： 线粒体凋亡通路中的重要凋亡刺激因子Smac与卵巢癌的发生发展及化疗耐药密切相关。我们通过临床标本检测发现Smac在卵巢良、恶性肿瘤组织中表达依次降低，且低分化、中晚期病例较高分化、早期病例的降低更为显著，二者具有统计学意义。本课题组依据Smac的N端AVPI功能序列，合成具有细胞穿透性的Smac多肽、Smac小分子。通过Smac多肽、Smac小分子以及外源性转染Smac基因三种途径共同研究发现Smac单独处理人卵巢癌细胞及耐药细胞，可抑制肿瘤细胞生长并刺激凋亡；分别联合紫杉醇或顺铂共同作用于人卵巢癌细胞及耐药细胞，可增强耐药细胞对化疗药物敏感性，降低耐药性。通过基因技术筛选多个相关凋亡基因，并分别对其下游基因进行检测，探讨Smac抑制肿瘤细胞生长、刺激凋亡及增强卵巢癌化疗敏感性的可能通路。体内动物试验进一步验证Smac高表达可在无显著毒副作用的基础上，有效增强卵巢癌化疗敏感性。本课题为卵巢癌化疗耐药或复发卵巢癌患者的治疗提供了实验依据，为开发、研制以Smac为靶点的新型靶向治疗药物奠定了数据基础。

中文关键词： 凋亡；耐药；卵巢癌；Smac多肽；类Smac小分子

英文摘要： Smac, as an important apoptotic stimulus in mitochondrial apoptosis pathway, has a relationship with ovarian cancer progression and chemotherapy resistance closely. we observed that Smac expression declined orderedly by benign and malignant ovarian tissue. And the expression of advanced cancer cases with poor differentiation was also significantly lower than that of early cases with well differentiated, both statistically significant. According to the N-terminal function sequences (AVPI) of Smac/DIABLO，we synthesized Smac polypeptide and Smac mimic molecule with cell membrane penetrability. We enhanced the intracellular expression of Smac/DIABLO in three different ways: transfected exogenous Smac plasmid, treated with Smac polypeptide and Smac mimic molecule. As a result, we found that over-expression of Smac alone inhibited the tumor cell growth and promoted apoptosis of human ovarian cancer cells. At the same time, over-expression of Smac improved the sensitivity of drug-resistant cells to chemotherapeutic drug, when dealing with paclitaxel or cisplatin together. In order to find the possible apoptosis pathway of Smac, we screened several apoptosis genes associated with Smac through gene technology, and detected their downstream genes respectively. It has been further confirmed that over-expression of Smac/DIABLO enhanced the sensitivity of ovarian cancer to chemotherapy significantly with no obvious side effects in vivo based on the experiments of animals. The results of this study provide experimental basis and data foundation for treatment of the drug-resistant or recurrent ovarian epithelial cancer cases by targeting Smac and providing targeting therapy of Smac.

英文关键词： ovarian cancer;drug resistence;apoptosis;Smac polypeptide;Smac mimic molecule;