Systematic identification of Celastrol-binding proteins reveals that Shoc2 is inhibited by Celastrol
Systematic identification of Celastrol-binding proteins reveals that Shoc2 is inhibited by Celastrol
复制标题
雷公藤红素结合蛋白的系统鉴定表明 Shoc2 被雷公藤红素抑制
DOI：
10.1042/bsr20181233
复制
发表时间：
2018-12-21
影响因子：
4
通讯作者：
Zhu Jiang-bo
中科院分区：
文献类型：
--
作者：
Huang Xiao-pei;Chen Ji-kuai;Zhu Jiang-bo
结直肠癌（CRC）是第三大最常诊断的癌症。雷公藤红素在多种癌症中表现出抗肿瘤活性。然而，Celastrol对人类结直肠癌的作用及其机制仍有待阐明。本研究旨在通过体外和体内两种方法阐明雷公藤红素的抗肿瘤作用，并利用蛋白芯片技术探讨其作用机制。我们证明Celastrol有效抑制SW480 CRC细胞增殖。两周的雷公藤红素灌胃可明显抑制裸鼠异种移植物的生长。在蛋白微阵列实验中，共鉴定出69个候选蛋白，其中富集程度最高的蛋白Shoc2，是一种通过ERK通路调节细胞运动和转移的支架蛋白。Celastrol在细胞系和异种移植瘤中显著抑制ERK1/2磷酸化。使用shoc2sirna下调Shoc2表达也抑制ERK1/2磷酸化。此外，下调Shoc2表达还能显著抑制肿瘤细胞的增殖、集落形成和迁移功能。此外，C57雄性小鼠灌胃对雷公藤红素的LD0≥80 mg/kg。综上所述，我们揭示了Celastrol的抗crc功能，并首次证实其通过与Shoc2结合抑制ERK1/2通路。
Colorectal cancer (CRC) is the third most commonly diagnosed cancer. Celastrol exhibits anti-tumor activities in a variety of cancers. However, the effect of Celastrol on human CRC and the underlying mechanisms still need to be elucidated. The present study aimed to use in vitro and in vivo methods to clarify the anti-tumor effect of Celastrol and use protein microarrays to explore its mechanisms. We demonstrated that Celastrol effectively inhibited SW480 CRC cell proliferation. Two weeks of Celastrol gavage significantly inhibited the growth of xenografts in nude mice. A total of 69 candidate proteins were identified in the protein microarray experiment, including the most highly enriched protein Shoc2, which is a scaffold protein that modulates cell motility and metastasis through the ERK pathway. Celastrol significantly inhibited ERK1/2 phosphorylation in cell lines and xenograft tumors. Down-regulation of Shoc2 expression using Shoc2 siRNA also inhibited ERK1/2 phosphorylation. Furthermore, down-regulation of Shoc2 expression also significantly inhibited proliferation, colony formation, and migration functions of tumor cells. In addition, the LD0 of Celastrol by gavage is equal or more than 80 mg/kg in C57 male mice. In summary, we unraveled the anti-CRC function of Celastrol and confirmed for the first time that it inhibited the ERK1/2 pathway through binding to Shoc2.