Colon cancer cell treatment with rose bengal generates a protective immune response via immunogenic cell death.
Colon cancer cell treatment with rose bengal generates a protective immune response via immunogenic cell death.
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DOI：
10.1038/cddis.2016.473
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发表时间：
2017-02-02
影响因子：
9
通讯作者：
Maker AV
中科院分区：
文献类型：
--
作者：
Qin J;Kunda N;Qiao G;Calata JF;Pardiwala K;Prabhakar BS;Maker AV
需要免疫治疗方法来管理晚期胃肠道恶性肿瘤患者；然而，激发肿瘤特异性免疫反应的机制仍有待阐明。玫瑰红（RB）在低浓度下对恶性细胞有毒，并可能诱导损伤相关的分子模式；因此，我们研究了它作为结肠癌免疫调节剂的潜力。小鼠和人结肠癌系用RB（10%盐水/PV-10）处理，进行细胞周期、细胞死亡和凋亡测定。用western blot、ELISA和流式细胞术评估损伤相关分子模式。在具有免疫功能的结肠癌小鼠模型中，我们证明了RB治疗后肿瘤消退，并且RB通过G2/M生长阻滞和主要坏死诱导结肠癌细胞死亡。rb处理的结肠癌细胞表现出不同的免疫原性细胞死亡（ICD）特征，包括细胞表面钙调蛋白和热休克蛋白90的表达增强，细胞内ATP减少，HMGB1释放。为了确认ICD表型，我们给免疫功能正常的动物接种了经RB处理的同基因结肠癌细胞。经rb处理的肿瘤可作为疫苗，抵抗同样的CT26结肠癌肿瘤细胞的后续攻击，用体外rb处理的细胞接种后，结肠癌细胞接种后肿瘤生长较慢，而非同基因的非CT26癌细胞接种后肿瘤生长较慢，表明具有特异性的抗肿瘤免疫应答。综上所述，RB作为ICD的诱导剂，有助于增强结直肠癌的特异性抗肿瘤免疫。
Immunotherapeutic approaches to manage patients with advanced gastrointestinal malignancies are desired; however, mechanisms to incite tumor-specific immune responses remain to be elucidated. Rose bengal (RB) is toxic at low concentrations to malignant cells and may induce damage-associated molecular patterns; therefore, we investigated its potential as an immunomodulator in colon cancer. Murine and human colon cancer lines were treated with RB (10% in saline/PV-10) for cell cycle, cell death, and apoptosis assays. Damage-associated molecular patterns were assessed with western blot, ELISA, and flow cytometry. In an immunocompetent murine model of colon cancer, we demonstrate that tumors regress upon RB treatment, and that RB induces cell death in colon cancer cells through G2/M growth arrest and predominantly necrosis. RB-treated colon cancer cells expressed distinct hallmarks of immunogenic cell death (ICD), including enhanced expression of calreticulin and heat-shock protein 90 on the cell surface, a decrease in intracellular ATP, and the release of HMGB1. To confirm the ICD phenotype, we vaccinated immunocompetent animals with syngeneic colon cancer cells treated with RB. RB-treated tumors served as a vaccine against subsequent challenge with the same CT26 colon cancer tumor cells, and vaccination with in vitro RB-treated cells resulted in slower tumor growth following inoculation with colon cancer cells, but not with syngeneic non-CT26 cancer cells, suggesting a specific antitumor immune response. In conclusion, RB serves as an inducer of ICD that contributes to enhanced specific antitumor immunity in colorectal cancer.