精子蛋白17单克隆抗体-多柔比星免疫偶联物体内外靶向抗卵巢癌活性研究  

Targeted antitumor activity of anti-Sp17 mAb-doxorubicin conjugates for ovarian cancer in vitro and in vivo

作　　者：曹王丽[1] 宋佳希[1] 李芳秋[1] 贾煊[1] 史利宁[1] 

机构地区：[1]南京军区南京总医院解放军临床检验医学研究所,江苏南京210002

出　　处：《中华肿瘤防治杂志》2012年第22期1685-1689,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81101978);国家自然科学基金(30670599);南京军区医学科学技术“十一五”研究计划重点项目(06Z43)

摘　　要：目的:将抗精子蛋白17(anti-Sp17)单克隆抗体(3C12)与多柔比星(DOX)偶联,并考察偶联物的体内外抗卵巢癌作用。方法:以二琥珀酰亚胺基酒石酸盐(DST)为交联剂制备3C12-DOX偶联物;免疫荧光法检测3C12-DOX的内化作用;MTT法测定3C12-DOX对Sp17+卵巢癌细胞的杀伤作用,流式细胞仪检测细胞的凋亡率。建立高表达Sp17荷瘤裸鼠模型,分为5组给药,分别经尾静脉注入生理盐水、3C12、S2D11-DOX、3C12-DOX和DOX,定期测量肿瘤体积和裸鼠体质量;实验结束时检测血液AST、LDH和CK-MB,检测心肌组织MDA和SOD,比较3C12-DOX和DOX对心脏的毒性。各组肿瘤称取质量,并检测肿瘤组织中细胞的凋亡率。结果:Sp17单克隆抗体(3C12)与DOX成功偶联,形成的3C12-DOX偶联物能够与Sp17+的细胞结合,并内化至细胞内;MTT结果显示,3C12-DOX偶联物能明显杀伤SK-OV3和HO8910细胞,加速细胞凋亡。在体实验中,注射3C12-DOX偶联物组的肿瘤体积明显小于其他对照组(DOX组除外)。而与DOX组对比,3C12-DOX毒副作用明显降低。结论:3C12-DOX是潜在的靶向治疗卵巢癌的药物。OBJECTIVE：To investigate whether a novel immunoconjugate containing anti-Sp17 monoclonal antibody （clone 3C12） linked to the chemotherapeutic agent doxorubicin [（DOX） Adriamycin] had antitumor activity against ovarian cancer cell lines and tumor models. METHODS： DOX was conjugated to 3C12 using a linker, and the specificity of 3C12-DOX was examined in SpiT-positive SKOV3 and Spl7-negative COC2 ovarian cancer ceils using cell-based ELISA and internalization assay. In vitro, the cytotoxicity of 3C12-DOX was determined by MTT and flow cytometry assay. In vi vo,its therapeutic effectiveness was evaluated in immunodeficient mice bearing SKOV3 cells. Five groups of tumor bearing mice were intravenously injected with normal saline, 3C12, S2Dll-DOX, 3C12-DOX or DOX respectively. Mice were weighed daily and tumors were measured twice weekly. AST, LDH, CK-MB, MDA and SOD were measured after mice died. Each excised tumors were weighed. Apoptosis of tumor cells was detected by flow cytometry assay. RESULTS： The 3C12-DOX immunoconjugate specifically bound to Sp17-positive SKOV3 but not to Sp17-negative cells and was internalized by SKOV3 cells. Treatment with 3C12-DOX decreased the viability of SKOV3 cells in an Spl7-specific manner. In vivo,3C12-DOX treatment,compared with control treatment, caused regression of established tumors of the mice with SKOV3 cells. The antitumor effects of 3C12-DOX were significantly associated with the induction of apoptosis in tumor cells. In addition, 3C12-DOX showed no observable adverse effects or toxicity when compared with DOX alone in mice bearing ovarian tumor xenografts. CONCLUSION： 3C12-DOX may be a potential antibodv-drug conjugate for clinical develooment.

关 键 词：卵巢肿瘤 精子蛋白17 多柔比星 靶向治疗 

分 类 号：R737.31[医药卫生—肿瘤]