Hu Fabiao, Pan Daxia, Zheng Wenyun, Yan Ting, He Xiujuan, Ren Fuzheng, Lu Yiming, Ma Xingyuan
State Key Laboratory of Bioreactor Engineering, and School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
Shanghai Key Laboratory of New Drug Design, and School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
Oncotarget. 2017 Dec 1;8(69):113687-113700. doi: 10.18632/oncotarget.22823. eCollection 2017 Dec 26.
Survivin was the smallest member of the IAP family, which was over expressed in many different cancers, and considered to be a promising hot target for cancer therapy, and our previous study demonstrated that multiple dominant negative mutants from full-length survivin could have many complex effects on cancer cells, such as cell cycle, apoptosis, and autophagy. But it was not yet known what role the two main domains played in those functions, which would be very important for the design of targeted anticancer drugs and for the interpretation of their molecular mechanisms. In this study, based on preparation the two parts (BIR domain and CC domain) of survivin by genetic engineering and cell characterization assay, we discovered that BIR (T34A)-domain peptide could inhibit Bcap-37 cells growth in a dose- and time-dependent manner, increase the proportion of G2/M phase, and induce caspase-dependent apoptosis via the mitochondrial pathway. While CC (T117A)-domain peptide increased the proportion of S-phase cells and increased the level of the autophagy marker protein LC3B significantly. These further experiments confirmed that TAT-BIR (T34A) peptide could be used to inhibit cell proliferation, promote apoptosis, and block mitosis, and TAT-CC (T117A) peptide showed mainly to promote autophagy, process of DNA replication, and mitosis to breast cancer cells. This research will lay the foundation for interpreting the multifunction mechanism of survivin in cell fates, further make senses in developing the anticancer drugs targeting it precisely and efficiently.
Survivin是凋亡抑制蛋白(IAP)家族中最小的成员,在多种不同癌症中过度表达,被认为是癌症治疗中一个很有前景的热门靶点。我们之前的研究表明,全长Survivin的多个显性负性突变体可能对癌细胞有许多复杂的影响,如细胞周期、凋亡和自噬。但尚不清楚这两个主要结构域在这些功能中发挥了什么作用,这对于靶向抗癌药物的设计及其分子机制的阐释非常重要。在本研究中,通过基因工程制备Survivin的两个部分(杆状病毒IAP重复序列(BIR)结构域和卷曲螺旋(CC)结构域)并进行细胞特性分析,我们发现BIR(T34A)结构域肽可以剂量和时间依赖性方式抑制Bcap-37细胞生长,增加G2/M期比例,并通过线粒体途径诱导半胱天冬酶依赖性凋亡。而CC(T117A)结构域肽增加了S期细胞比例,并显著提高了自噬标记蛋白微管相关蛋白1轻链3β(LC3B)的水平。这些进一步的实验证实,穿膜肽-TAT-BIR(T34A)肽可用于抑制细胞增殖、促进凋亡和阻断有丝分裂,而穿膜肽-TAT-CC(T117A)肽主要显示为促进乳腺癌细胞的自噬、DNA复制过程和有丝分裂。本研究将为阐释Survivin在细胞命运中的多功能机制奠定基础,进一步为精准高效地开发靶向它的抗癌药物提供依据。