NFAT3在黑素瘤中的功能及其作为治疗靶点的研究

发布时间：2018-02-16 16:11

本文关键词： 黑素瘤 NFAT3 FK506 FK520 增殖迁移　出处：《深圳大学》2017年硕士论文　论文类型：学位论文

【摘要】：黑素瘤(Melanoma)是皮肤癌中恶性程度最高的一种亚型,拥有极高的致死率,接近五分之四的皮肤癌致死病例均由黑素瘤引起。2012年全世界共有203200例黑素瘤患者,造成55000人死亡。目前治疗黑素瘤的主要手段为放化疗或手术治疗,但是常规的治疗手段所取得的效果有限且具有极大的副作用,因此,研究发现黑素瘤发生发展中的关键靶标,进而寻找特异性药物,成为当今黑素瘤治疗的重要策略。活化T细胞核因子(Nuclear factor of activated T cells,NFAT)家族是一类在免疫反应中能够调节促炎细胞因子和其它基因表达的转录因子。除免疫细胞外,如今已经被发现在肌肉、骨骼、神经元、内脏、皮肤中均有表达并与癌症密切相关。相比较于其它NFAT家族成员,NFAT3在癌症中的研究较少而且结论不一致。目前NFAT3在黑素瘤中的功能尚属空白。他克莫斯(FK506)和子囊霉素(FK520)化学结构相似,均为临床上常见的抗器官移植后的免疫排斥反应用药。它们能够通过抑制钙调磷酸酶(CaN)的活性,降低NFAT蛋白去磷酸化的程度,抑制NFAT蛋白入核从而发挥免疫抑制作用。FK506目前被作为一种潜在的癌症治疗药物被应用于多种癌症的临床前研究,如膀胱癌、前列腺癌、乳腺癌等。FK506作用于黑素瘤细胞的黑素原生成也见于报道。但目前尚无FK520与黑素瘤治疗相关的研究,且NFAT3与FK506、FK520的相关性不明。在前期NFAT3相关研究的基础上,本研究检测了NFAT3在黑素瘤细胞系的表达,选取NFAT3高表达的黑素瘤细胞A375、SK-MEL-28构建NFAT3敲低稳定株并且进行功能实验。我们发现敲低NFAT3的表达能抑制黑素瘤细胞的增殖和迁移,结果说明NFAT3作为一种促癌因子在黑素瘤中起作用。随后使用FK506、FK520处理黑素瘤A375、SK-MEL-28细胞,结果发现FK506、FK520能抑制黑素瘤细胞的增殖和迁移;进一步实验发现FK506、FK520抑制NFAT3的核定位而影响NFAT3的核质分布;最后通过裸鼠皮下荷瘤实验显示FK506、FK520在活体水平上对黑素瘤生长的抑制作用。综上,可以得出结论,NFAT3在黑素瘤进程中是一个促癌基因并且是一个潜在的黑素瘤治疗靶点,而FK506和FK520能够通过抑制NFAT3的功能从而阻止黑素瘤的生长和迁移。
[Abstract]:Melanoma (melanoma) is the most malignant subtype of skin cancer, with a high mortality rate. Nearly 4/5 cases of skin cancer deaths are caused by melanoma. In 2012, there were 203200 cases of melanoma in the world. At present, the main treatment for melanoma is radiotherapy, chemotherapy or surgery, but the effect of conventional treatment is limited and has great side effects. Therefore, the study found that melanoma is a key target in the development of melanoma. And then looking for specific drugs, Nuclear factor of activated T cells (NFATs) family is a class of transcription factors that regulate the expression of proinflammatory cytokines and other genes in the immune response. Now it's been found in muscles, bones, neurons, viscera, Compared with other members of the NFAT family, NFAT3 is less studied in cancer and the results are inconsistent. At present, the function of NFAT3 in melanoma is still blank. Tacrolimus FK506) and ascomycetes. Its chemical structure is similar to that of FK520. They can reduce the degree of dephosphorylation of NFAT protein by inhibiting the activity of calmodulin. Inhibiting the entry of NFAT protein to play an immunosuppressive effect. FK506 is currently being used as a potential cancer treatment drug in preclinical studies of many cancers, such as bladder cancer, prostate cancer, The expression of melanin in melanoma cells induced by breast cancer. FK506 is also reported. However, there is no research on the relationship between FK520 and melanoma therapy, and the correlation between NFAT3 and FK506FK520 is not clear. In this study, we detected the expression of NFAT3 in melanoma cell line, and constructed the stable NFAT3 knockdown cell line A375 SK-MEL-28 with high NFAT3 expression. We found that low NFAT3 expression could inhibit the proliferation and migration of melanoma cells. The results showed that NFAT3 acts as a carcinogenic factor in melanoma, and then treated with FK506 and FK520 in melanoma cell line A375 SK-MEL-28. The results showed that FK506 and FK520 could inhibit the proliferation and migration of melanoma cells. It was further found that FK506FK520 inhibited the nuclear localization of NFAT3 and affected the distribution of nuclear and cytoplasm of NFAT3. Finally, it was shown that FK506FK520 inhibited the growth of melanoma in vivo by subcutaneous tumor implantation in nude mice. It can be concluded that NFAT3 is a carcinogenic gene and a potential target for melanoma therapy in the process of melanoma, while FK506 and FK520 can inhibit the growth and migration of melanoma by inhibiting the function of NFAT3.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.5

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