Genistein靶向多肽偶联物的制备及体外抑瘤活性研究  

Preparation of genistein-targeting peptide conjugate and its anti-tumor activity in vitro

作　　者：项贵明[1] 粟永萍[1] 程天民[1] 艾国平[1] 

机构地区：[1]第三军医大学军事预防医学院全军复合伤研究所创伤,烧伤与复合伤国家重点实验室,重庆400038

出　　处：《中华肿瘤防治杂志》2006年第21期1614-1616,1626,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：重庆市院士专项基金(2004BC5006)

摘　　要：目的:制备Genistein靶向多肽偶联物(GEN-P),增强Genistein的抑瘤活性。方法:采用固相法设计、合成能够与组织因子(TF)特异性结合的配体分子-FHS001,凝血酶原时间(PT)法测定其凝血活性,荧光显微镜直接检测其与高表达TF的人乳腺癌细胞株MCF-7的结合。通过异型双功能交联剂Sul-fo-SANPAH将FHS001与Genistein交联,制备偶联物GEN-P;MTT法检测其对MCF-7的杀伤作用。结果:合成的FHS001多肽能够与高表达TF的MCF-7细胞特异性结合且无明显的凝血活性。GEN-P偶联物在0.2～200.0μmol/L的浓度范围内对MCF-7细胞均有不同程度的增殖抑制作用,作用6h后抑制率为16.3%～83.1%,且呈一定的剂量依赖性。GEN-P和Genistein对MCF-7细胞的IC50分别为2.0和120μmol/L,GEN-P效果明显优于游离药物Genistein。结论:与FHS001偶联后Genistein的抑瘤活性明显增强,有希望进一步应用于癌症的治疗。ORJECTIVE：To enhance the activity of genistein against cancer cells by coupling it to a targeting peptide. METHODs： Peptide FHS001 was designed and synthesized with solid phase peptide synthesis. Its clotting activity was measured by the prothrombin time （PT） assays. Then specific binding activity of FHS001 was detected by fluorescence microscopy using human breast cancer cells MCF-7 on which TF was highly expressed. GEN-P conjugate was prepared by using heterobifunctional crosslinker sulfosuccinimidyl 6- （4′-azido-2′-nitrophenylamino） hexanoate （ Sulfo-SANPAH ）. The cytotoxicity of GEN-P was evaluated by MTT assay. RESLUTS： The FHS001 peptide designed by us had no clotting activity and had specific binding activity to TF. GEN-P was achieved by photochemical conjugation. In antiproliferation assays, the inhibition rates of treatment with 0. 2-200. 0 μmol/L GEN-P were from 16. 3% to 83. 1 %. The IC50 for GEN-P and genistein against MCF-7 cells were 2. 0 μmol/L and 120 μmol/L respectively. GEN-P inhibited MCF-7 cells in a dose-dependent manner and was at least 50 times more effective than genistein alone. CONCLUSION.. The conjugating genistein to targeting peptide FHS001 enhances its cytotoxicity. This may constitute a novel approach for carcinoma therapy.

关 键 词：染料木黄酮/药理学 乳腺肿瘤/病理学 肿瘤细胞 培养的/药物作用 

分 类 号：R737.9[医药卫生—肿瘤] R730.53[医药卫生—临床医学]