乌贼墨寡肽EPSI-1的制备与抗前列腺癌机制研究

前列腺癌（PCa）是全球最常见恶性肿瘤之一，一旦发生或转移，治疗效果较差，且肿瘤长期化疗损害重要脏器，也会产生肿瘤耐药。因此，寻找低毒、高效新型抗癌药物势在必行。在研究PCa发生、转移及相关基因表达基础上，近5年来，又从乌贼墨中经酶解法首次得到5种不同分子量乌贼墨多肽（EPSI），其中分子量小于1KDa的EPSI-1具有显著诱导PCa细胞凋亡和基因调控作用。采用GE、GC、HPLC、CD和NMR等对EPSI-1进行分离纯化和结构表征。对其纯化物进行抗肿瘤机制研究，采用PCa细胞株和荷瘤小鼠模型进行体内外试验，阐明PCa的p53、nm23H1、VEGF-C表达以及MVD、MLC变化，明确海洋药物筛选和制备的关键技术，评价其抗PCa作用机制。本研究将制备一种新型抗PCa海洋药物前身物，为一种具有自主知识产权的抗肿瘤海洋肽类药物的研发奠定基础。因此，具有重要的学术价值和较好的应用前景。

Prostate cancer is one of the most common malignant tumors all over the world. Most prostate cancer are low growing. However, there are cases of aggressive prostate cancers. Toxicity, drug resistance and limited transient benefits in patients are the main problems associated with standard chemotherapeutic regimens, and new drugs are therefore needed to fight this devastating disease. In recent five years, we get EPSI of five different molecular weights defined as EPSI-1, EPSI-2, EPSI-3, EPSI-4 and EPSI-5,respectively. EPSI-1 whose molecular weight is under 1KDa possessed high anti-cancer properties including cell apoptosis and regulated the gen expression. EPSI-1 is isolated by ultrafiltration, purified on G-25 gel filtration and HPLC and demonstrated the purity of EPSI-1 by HPLC.Its peptide sequence analysis is detected. What's more, the structure of EPSI-1 is identified by NMR, GC and GE. The anti-tumor effect and mechanism of purified EPSI-1 in vitro and in vivo are investigated. The hormone-independent DU-145 and PC-3 are used to study the anti-tumor effect and mechanism of purified EPSI-1 in vitro. Concentration-dependent responses of the effect of EPSI-1on cell proliferation is examined using CCK-8 assay. AO/EB staining and FCM are employed for observation of apoptosis on DU-145 and PC-3 cells after treating with EPSI-1.WB, FQ-PCR and ISH are used to analyze the expression of Bcl-2,Caspase-3, Caspase-9,p53,mm23H1and VEGF-C in PC-3 and DU-145 cells treated with EPSI-1.The nude mouse model of human prostate cancer is established and its expression of p53,mm23H1,VEGF-C and MVD and the number of MLC are studied. From the above experiment ,the key technology of marine drug selection and preparation can be cleared and defined .This study will prepared a new anti-prostate cancer predecessor of marine drug which can lay a foundation for the development of peptide drug with independent intellectual property. Therefor ,this study possesses important academic values and a good perspective of application.