抗肿瘤药物的代谢产物N-乙酰基-S-（对氯苯甲酰基）半胱氨酸抑制恶性黑色素瘤生长的作用和机理研究

项目名称： 抗肿瘤药物的代谢产物N-乙酰基-S-（对氯苯甲酰基）半胱氨酸抑制恶性黑色素瘤生长的作用和机理研究

项目编号： No.81202553

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 药物学、药理学

项目作者： 陈巍

作者单位： 浙江省肿瘤医院

项目金额： 23万元

中文摘要： NACC是抗肿瘤药物磺氯苯脲的主要代谢产物，其蛋白结合较磺氯苯脲大幅降低。前期研究表明：NACC对体外培养人恶性黑色素瘤细胞、小鼠皮肤癌模型均有显著的抑制作用，而对正常猴肾细胞无明显毒性。NACC对细胞周期无影响，但能诱导细胞凋亡；能抑制谷氧还蛋白和硫氧还蛋白的活性和黑色素瘤细胞的转移。 本项目拟在前期研究的基础上，采用细胞培养和裸鼠肿瘤移植实验，进一步评价NACC对黑色素瘤细胞的抑制作用。采用Annexin-V/PI双染法考察NACC对黑色素瘤细胞凋亡的影响；采用荧光光度法、酶动力学分析、流式细胞术、ELISA、Western blot、RT-PCR等方法考察NACC对肿瘤细胞胞浆钙离子浓度、线粒体跨膜电位，肿瘤细胞谷氧还蛋白、硫氧还蛋白和硫氧还蛋白还原酶活性，促凋亡蛋白表达的影响；利用细胞和裸鼠模型考察NACC对黑色素瘤细胞侵袭和转移的影响；以明确NACC抑制黑色素瘤细胞生长的机理。

中文关键词： 恶性黑色素瘤；NACC；细胞凋亡；迁移；硫氧还蛋白还原酶

英文摘要： N-Acetyl-S-(p-chlorophenylcarbamoyl)cysteine (NACC) is one of the major metabolites in rats of anticancer drug-sulofenur developed by Elli Lilly research laboratories. Sulofenur has exhibited decent and broad anticancer activity against human solid tumors in clinical trials. However, due to its high protein binding, a high dosage of sulofenur was required for clinical responses. As a consequence, its side effect-anemia was observed in clinical trials. Our earlier study showed NACC had much lower protein binding compared to sulofenur and it had anticancer activity against, especially, human melanoma, also UV-induced skin cancer in hairless mouse model. It induces apoptosis in melanoma cells but has no effect on cell cycle. It is able to inhibit thioredoxin and glutaredoxin activity. It inhibits metastasis of melanoma cells in vitro. It is known that sulofenur is able to elevate cytosolic Ca2+ concentration and decrease mitochondrial membrane potential by which apoptosis could be induced. It is very possible that the mechanism of action of NACC will be elevation of cytosolic Ca2+ concentration and then induction of apoptosis in cancer cells through mitochondrial pathway. In this study, the anticancer activity of NACC against melanoma will be further evaluated in vitro and in vivo by using cultured huma

英文关键词： Melanoma；NACC；Apoptosis；Migration；TrxR