乳源免疫六肽抗卵巢癌及其机制的研究

项目名称： 乳源免疫六肽抗卵巢癌及其机制的研究

项目编号： No.30872992

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 秦宜德

作者单位： 安徽医科大学

项目金额： 30万元

中文摘要： 卵巢癌是妇科最为严重的恶性肿瘤。乳源免疫六肽（PGPIPN）来源于牛乳中β#37226;蛋白的f63-68。本项目研究PGPIPN抗卵巢癌功能及其机理，为开发PGPIPN作为新的肽类抗肿瘤药物提供基础，并为卵巢癌提供新的治疗手段。通过体内外两种方式，利用细胞培养及荷瘤动物模型等探索PGPIPN对卵巢癌细胞和肿瘤生长的影响；并通过动物模型，研究PGPIPN对其免疫和内分泌的影响。结果表明，在体外，PGPIPN能显著抑制人卵巢癌细胞（SKOV3）的增殖并诱导其凋亡（最高凋亡率30.73%），其效果与作用时间和剂量相关。通过荷SKOV3裸鼠模型实验，表明PGPIPN能显著抑制肿瘤的生长，最高抑制率68.46%，其效果优于传统抗癌药物5氟尿嘧啶（抑制率41.54%），同时能显著提高荷瘤动物的脾指数和健康水平。PGPIPN能提高淋巴细胞的转化和机体的免疫功能，PGPIPN也能降低荷瘤动物的IGF-1的水平，从而影响其内分泌功能。以管家基因作参照，PGPIPN能抑制bcl-2等凋亡抑制基因表达，而促进bax等凋亡相关基因表达。PGPIPN作用位点在细胞膜上，可以通过卵巢癌细胞膜上受体而直接影响卵巢癌细胞。

中文关键词： PGPIPN；SKOV3；人卵巢癌；抗癌；凋亡

英文摘要： Ovarian cancer is the most serious gynecological malignant tumor, and the mortality rates being the biggest in gynecologic tumors. Immunomodulatory hexapeptide (PGPIPN) is derived from bovine beta-casein f63-68. This project studied anti-ovarian cancer of PGPIPN and its mechanism, which could provide a basis for developping PGPIPN as a new peptide anticancer drugs and a new therapeutic method for ovarian cancer. The effects of PGPIPN on ovarian cancer cells and tumor growth were studied by cell culture in vitro and model of xenograft nude mice taking human ovarian cancer (SKOV3). Meanwhile, we also studied the impact of PGPIPN on immune and endocrine of body by animal model. Experimental methods in the project including electron microscopy, DNA fragmentation detection, immunohistochemistry, flow cytometry analysis, cDNA chip, fluorescence localization, affinity chromatography, mass spectrometry. The results showed that PGPIPN can significantly inhibit the proliferation of human ovarian cancer cell lines (SKOV3) and induced its apoptosis in vitro (P <0.05 or P <0.01), and the effect varied with time and dosage dependent manner. Also, the morphological changes of SKOV3 apoptosis induced by PGPIPN had been observed. According to FCM analysis, the highest apoptosis rated at 30.73% appeared in 3×-4 g/L. PGPIPN can significantly suppress the growth of xenograft ovarian tumors in vivo in a dose-dependent manner (P <0.05 or P <0.01). The anti-ovarian carcinoma effect (68.46% inhibitory rate) of high dose PGPIPN exceeded that (41.54% inhibitory rate) of traditional anti-ovarian drug, 5-fluorouracil. The result also suggested PGPIPN could promote spleen hyperplasia of xenograft nude mice, and presented the better state of health of xenograft nude mice. The PGPIPN can improve the lymphocyte transformation and immune function of body. Moreover, PGPIPN treatment leads to significantly down-regulation of serum IGF-1 in the xenograft mice, thus affecting body's endocrine. Taking housekeeping genes as a reference PGPIPN can inhibit the expression of apoptosis suppressor gene bcl-2, while promoting the expression of apoptosis-related genes bax (P <0.05 or P <0.01). The PGPIPN receptor was located in the cell membrane, which could directly affect ovarian cancer by the receptor. These findings reveal that PGPIPN attenuates the tumorigenesis of human ovarian cancer cells through multiple mechanisms, and suggest that PGPIPN is a promising drug for the treatment of ovarian cancer.

英文关键词： PGPIPN; SKOV3; human ovarian carcinoma anti-cancer; apoptosis