海洋中药毛蚶抗肿瘤活性物质基础及其作用机制研究

Arca subcrenata Lischke is one kind of traditional Chinese medicines from marine, which has many efficacies such as blood enriching, spleen-stomach warming and stomach invigorating. Our previous research indicated that the P2 component of A.subcrenata could induce the apoptosis and cycle arrest of HT-29 and HeLa cells in a dose-dependent manner.The current study aims to systemically isolate and purify the P2 component, elucidate the material basis for its efficacies, and explore its antitumor mechanisms. The content of the proposed study includes the isolation and purification of the bioactive polypeptides by chromatography, preparative HPLC and electrophoresis under the bioassay-guidance of antitumor activity,the structural determination of the bioactive polypeptides by spectroscopic methods and combined techniques,the evaluation of the in vitro antitumor activity of the bioactive polypeptides by MTT method and their in vivo antitumor activity with human tumor xenograft model in BALB/c mice, as well as the exploration of the antitumor mechanisms by molecular biology methods. This study marks the first time to systemically isolate and identify the bioactive polypeptides from A. subcrenata,evaluate their antitumor activity and explore the underlying mechanisms.The research findings will fill the gaps on the basic investigation of bioactive polypeptides from A. subcrenata, and provide scientific basis for research and development of novel drugs from marine organisms over which our country would possess intellectual property rights.

毛蚶为传统海洋中药，具有补血、温中、健胃等功效。我们前期研究提示：其P2组分可剂量依赖性诱导HT-29和HeLa细胞凋亡与周期阻滞。本课题拟在国家重大专项工作基础上，对多肽有效部位P2进行系统分离纯化，阐明其抗肿瘤作用药效物质基础，探究其抗肿瘤作用机制。内容包括：在抗肿瘤活性追踪指导下，利用各种柱层析、制备性HPLC、电泳等技术，分离纯化P2中的活性多肽化合物；利用各种波谱方法及多种联用技术鉴定其分子结构；采用MTT法评价多肽化合物的体外抗肿瘤活性；利用裸鼠人癌移植瘤模型评价其体内抗肿瘤作用；利用分子生物学手段，研究活性多肽化合物的作用靶点，探索其抗肿瘤作用机制。此项目系国内外首次对毛蚶P2组分中活性多肽化合物进行系统的分离鉴定，评价其抗肿瘤作用并探究作用机制。研究结果既可填补海洋中药毛蚶P2中活性多肽化合物基础研究空白，也可为研究和开发具有我国自主知识产权的海洋中药新药提供科学依据。

毛蚶为传统海洋中药，具有补血、温中、健胃等功效。我们前期研究提示：其P2组分可剂量依赖性诱导HT29和HeLa细胞凋亡与周期阻滞。本课题在前期工作基础上，对P2有效部位进行了系统分离纯化，研究其抗肿瘤作用的有效物质基础，探究其抗肿瘤作用的分子机制。取得如下研究成果：（1）在活性追踪指导下，采用各种层析色谱、制备HPLC、电泳等技术，分离纯化P2中的各种活性成分，得到了H3、H4等多种活性蛋白；（2）利用各种波谱方法及多种联用技术确定各蛋白化合物的化学结构；（3）采用MTT法评价有效部位及蛋白成分的体外抗肿瘤活性；（4）利用裸鼠人癌移植瘤模型评价单体的体内抗肿瘤作用；（5）利用分子生物学手段，研究有效单体的抗肿瘤作用靶点，探索其抗肿瘤作用机制。此项目系国内外首次对毛蚶P2有效部位进行系统的抗肿瘤活性成分研究，该研究结果既可为中药毛蚶有效部位/活性单体提供基础研究资料，也可为其进一步的创新药物研发提供科学依据。