青钱柳多糖CPP-1的精细结构及其抗肿瘤活性构效关系

Polysaccharide is an important biomolecules which continue the genic information.In recent years, it has been demonstrated that polysaccharide has significant antitumor activities both in vivo and in vitro. However,the molecular basis, mechanisms of these activities and the structure-activity relationship of polysaccharide are still unknown. Based on the previous work which has extracted and purified an antitumor polysaccharide of CPP-1 from Cyclocarya paliurus (Batal.) Iljinskaja, the present study is trying to analyze the physicochemical properties, molecular weight and chemical composition (including the monosaccharide, uronic acid and amine acid composition) of CPP-1 with modern instrumental analysis. In addition, the fine structure of CPP-1 is going to be studied with completely acid hydrolysis, X-ray, and circular dichroism (CD) to elucidate the backbone and side chain composition, the order of connection among sugar residues and the release of sugar chain. Furthermore, the antitumor effects of CPP-1 will be evaluated on the model of HeLa cell, and assessed by the expression of caspase-3 mRNA and caspase-3 protein. Thus，the mechanism of inducing cell apoptosis will be discussed at genetic and protein level with cell biology and molecular biology methods，and the relationship between structure and antitumor activity will be concluded. This study will provide important theory basis and technique supports for the research of structure-antitumor activity relationship for other plant polysaccharides.

多糖是一类重要的生物信息分子，是基因的信息延续，现已证明多糖具有显著的抗肿瘤生物活性，但其活性功能的分子基础、机理及构效关系尚不清楚。本项目以从我国特色资源青钱柳中分离纯化出的具有抗肿瘤活性的多糖CPP-1 为研究对象，采用红外光谱、激光拉曼光谱、色谱质谱联用、核磁共振谱等现代仪器分析方法，研究其理化性质，分子量、纯度，单糖、糖醛酸、氨基酸等精确组成，采用完全酸水解方法、X-射线和圆二色谱等研究其糖链精细结构如主链和支链的组成、糖残基间的连接顺序、连接方式及糖链释放等，从而解析其精细结构，并结合细胞生物学与分子生物学方法研究多糖对HeLa细胞凋亡、caspase-3 mRNA 及caspase-3 等蛋白表达的影响，从基因水平、蛋白水平探讨CPP-1 诱导HeLa细胞凋亡的作用机理，在此基础上探讨其抗肿瘤活性的构效关系。研究结果为植物多糖抗肿瘤活性构效关系的系统研究提供重要的方法学参考。

多糖是重要生物信息分子，是基因的信息延续，在基因组学和蛋白质组学发展同时，糖化学以及糖生物学研究也备受重视。多糖具有显著的抗肿瘤生物活性已为人所知，但其活性功能的机理及构效关系尚不清楚。. 项目以从江西特色资源青钱柳中分离纯化出的活性多糖为研究对象，采用食品化学与现代仪器分析方法，研究了青钱柳多糖的理化性质，分子量、纯度，单纯、糖醛酸、氨基酸等精确组成，分析了其糖链精细结构如主链和支链的组成、糖残基间的连接顺序等；研究 CPP 对 HeLa 细胞凋亡、Caspase-3 mRNA 及 Caspase-3 蛋白表达的影响，从基因水平、蛋白水平探讨 CPP 诱导 HeLa 细胞凋亡的作用机理，探讨其构效关系。. 项目达到预期目标，在国内外期刊上发表14篇（全部标注了基金资助，详见附件清单，其中SCI 11篇，IF大于4的SCI论文8篇，EI1篇），申请国家发明专利1项,受邀在国际学术报告上作学术报告2次，研究结果可为植物多糖抗肿瘤活性构效关系的系统研究提供参考。。