Department of Cellular Biology, Institute of Biology and Ecology, Faculty of Science, Pavol Jozef Šafárik University in Košice, Šrobárova 2, 041 54 Košice, Slovak Republic.
Department of Cellular Biology, Institute of Biology and Ecology, Faculty of Science, Pavol Jozef Šafárik University in Košice, Šrobárova 2, 041 54 Košice, Slovak Republic.
Toxicol In Vitro. 2020 Aug;66:104860. doi: 10.1016/j.tiv.2020.104860. Epub 2020 Apr 13.
The use of natural products as chemotherapeutic agents and tools for manipulation of apoptosis represent an attractive therapeutic concept. In this study, we investigated the anticancer activities of a combination of two natural compounds with different origin, hypericin (plant product) in its photoactive state and Manumycin A (yeast product) and explored the underlying mechanisms of their pro-apoptotic action using an oxaliplatin-resistant variant of human colon adenocarcinoma cell line HT-29-OxR as the experimental model. CCK-8 assay was performed to evaluate the cytotoxicity of the drugs. CalcuSyn software was used to identify the type of interaction between the two agents. BrdU incorporation assay and colony forming assay were performed to study the short- and long-term proliferation of cells. To evaluate the ability of the drug combination to induce apoptosis, PARP p85 fragment was detected using the ELISA method. Changes in apoptosis-related proteins were examined by immunoassays. Our results showed that a synergistic combination of photoactive hypericin and Manumycin A decreased viability, inhibited both short- and long-term cell proliferation, decreased levels of IAPs proteins (cIAP1, cIAP2, XIAP and survivin), induced an apoptotic PARP cleavage associated with decline in procaspase-3 level, promoted phagocytosis of cancer cells, and restored chemosensitivity to oxaliplatin.
天然产物作为化疗药物和调控细胞凋亡的工具的应用代表了一种有吸引力的治疗概念。在这项研究中，我们研究了两种具有不同来源的天然化合物组合的抗癌活性，即处于光激活状态的金丝桃素（植物产物）和马努菌素 A（酵母产物），并使用奥沙利铂耐药的人结肠腺癌细胞系 HT-29-OxR 的变体作为实验模型，探索了它们促凋亡作用的潜在机制。CCK-8 测定法用于评估药物的细胞毒性。CalcuSyn 软件用于识别两种药物之间的相互作用类型。BrdU 掺入测定法和集落形成测定法用于研究细胞的短期和长期增殖。为了评估药物组合诱导细胞凋亡的能力，使用 ELISA 方法检测 PARP p85 片段。通过免疫测定法检查与细胞凋亡相关的蛋白质的变化。我们的结果表明，光激活金丝桃素和马努菌素 A 的协同组合降低了细胞活力，抑制了短期和长期的细胞增殖，降低了 IAPs 蛋白（cIAP1、cIAP2、XIAP 和 survivin）的水平，诱导了与 procaspase-3 水平下降相关的凋亡 PARP 切割，促进了癌细胞的吞噬作用，并恢复了对奥沙利铂的化疗敏感性。
