Celastrol induces ROS-mediated apoptosis via directly targeting peroxiredoxin-2 in gastric cancer cells
Celastrol induces ROS-mediated apoptosis via directly targeting peroxiredoxin-2 in gastric cancer cells
复制标题
雷公藤红素通过直接靶向过氧化还蛋白-2诱导胃癌细胞ROS介导的细胞凋亡
DOI：
10.7150/thno.46728
复制
发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Liang, Guang
中科院分区：
文献类型：
--
作者：
Chen, Xi;Zhao, Ying;Liang, Guang
背景：据报道，活性氧簇(ROS)升高引起的氧化应激可诱导细胞凋亡，并可能提供靶向肿瘤细胞的手段。雷公藤红素是一种天然的生物活性化合物，最近被证明能增加ROS水平并导致癌细胞凋亡。然而，雷公藤红素的细胞毒作用机制尚不清楚，其直接的分子靶点也尚未确定。方法：采用蛋白质组芯片、表面等离子体共振、恒温滴定量热法和分子模拟等方法对雷公藤红素的分子靶点进行鉴定。通过结合和活性分析验证雷公藤红素与目标蛋白在无细胞和胃癌细胞裂解物中的相互作用。然后，我们评估了从胃癌患者获得的活检标本中的靶转录水平。在BALB/c nu/nu小鼠体内评价雷公藤红素的生长抑制和细胞毒活性。结果：雷公藤红素直接与抗氧化酶-2(Prdx2)结合，从而在分子和细胞水平上抑制其酶活性。雷公藤红素抑制Prdx2可增加细胞内ROS水平，并导致ROS依赖的内质网应激、线粒体功能障碍和胃癌细胞的凋亡。功能测试表明，雷公藤红素至少部分地通过靶向Prdx2来限制胃癌细胞。雷公藤红素对移植有胃癌细胞的小鼠的治疗也抑制了肿瘤的生长，与Prdx2抑制和ROS增加有关。对人类胃癌的分析也显示Prdx2水平升高，并与生存相关。结论：我们的研究发现了一种潜在的雷公藤红素相互作用蛋白Prdx2及其作用的ROS依赖机制。这些发现还强调Prdx2是治疗胃癌的潜在靶点。
Background: Oxidative stress from elevated reactive oxygen species (ROS) has been reported to induce cell apoptosis and may provide a means to target cancer cells. Celastrol is a natural bioactive compound that was recently shown to increase ROS levels and cause apoptosis in cancer cells. However, the underlying mechanism for this cytotoxic action remains unclear and direct molecular targets of Celastrol have not been identified.Methods: Proteome microarray, surface plasmon resonance, isothermal titration calorimetry and molecular simulation were used to identify the molecular target of Celastrol. Binding and activity assays were used to validate the interaction of Celastrol with target protein in cell-free and gastric cancer cell lysates. We then assessed target transcript levels in in biopsy specimens obtained from patients with gastric cancer. Gastric cancer growth-limiting and cytotoxic activity of Celastrol was evaluated in BALB/c nu/nu mice.Results: Our data show that Celastrol directly binds to an antioxidant enzyme, peroxiredoxin-2 (Prdx2), which then inhibits its enzyme activity at both molecular and cellular level. Inhibition of Prdx2 by Celastrol increased cellular ROS levels and led to ROS-dependent endoplasmic reticulum stress, mitochondrial dysfunction, and apoptosis in gastric cancer cells. Functional tests demonstrated that Celastrol limits gastric cancer cells, at least in part, through targeting Prdx2. Celastrol treatment of mice implanted with gastric cancer cells also inhibited tumor growth, associated with Prdx2 inhibition and increased ROS. Analysis of human gastric cancer also showed increased Prdx2 levels and correlation with survival.Conclusion: Our studies have uncovered a potential Celastrol-interacting protein Prdx2 and a ROS-dependent mechanism of its action. The findings also highlight Prdx2 as a potential target for the treatment of gastric cancer.