Toxicarioside G (TCG), a natural product isolated from Calotropis gigantea, has been found to exhibit potent anticancer effects. The present study aimed to investigate the effect of TCG on the SW480 colorectal cancer cell line and the role of autophagy and Yes1 associated transcriptional regulator (YAP) in the TCG‑mediated inhibition of cell proliferation and viability. Cell proliferation was detected using MTT, BrdU, colony formation and LDH release assays, while apoptosis was analyzed using flow cytometry and western blot analyses. Immunofluorescence and western blot analysis was used to determine TCG‑induced autophagy and YAP activation. Pharmacological inhibition and siRNA was used to investigate the role of autophagy and YAP in TCG‑mediated cell growth inhibition. The results revealed that TCG inhibited SW480 cell proliferation and viability, independent of apoptosis, and also induced autophagy. It was further demonstrated that TCG blocks autophagic flux, resulting in autophagy arrest in the SW480 cell line. The inhibition of autophagy restored the TCG‑mediated inhibition of cell proliferation and viability, suggesting that TCG may induce lethal autophagy arrest in the SW480 cell line. Furthermore, TCG induced YAP activation in the SW480 cell line. Inhibition of YAP activity enhanced the TCG‑mediated inhibition of cell proliferation and viability, suggesting that YAP may play a protective role in the TCG‑induced effects. In conclusion, the findings of the present study indicated that TCG may induce lethal autophagy arrest and activate YAP, which serves a protective role in the SW480 cell line. These results suggested that the combined targeting of TCG and YAP may represent a promising strategy for TCG‑mediated anticancer therapy.

中文翻译：

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YAP 激活减弱毒甙 G 诱导的 SW480 结直肠癌细胞中的致死性自噬停滞。

Toxicarioside G (TCG)，一种从Calotropis gigantea 中分离的天然产物, 已被发现具有强大的抗癌作用。本研究旨在研究 TCG 对 SW480 结直肠癌细胞系的影响以及自噬和 Yes1 相关转录调节因子 (YAP) 在 TCG 介导的细胞增殖和活力抑制中的作用。使用 MTT、BrdU、集落形成和 LDH 释放试验检测细胞增殖，而使用流式细胞术和蛋白质印迹分析分析细胞凋亡。免疫荧光和蛋白质印迹分析用于确定 TCG 诱导的自噬和 YAP 激活。药理抑制和 siRNA 用于研究自噬和 YAP 在 TCG 介导的细胞生长抑制中的作用。结果表明，TCG 抑制 SW480 细胞增殖和活力，独立于细胞凋亡，并且还诱导自噬。进一步证明 TCG 阻断自噬通量，导致 SW480 细胞系中的自噬停滞。自噬的抑制恢复了 TCG 介导的细胞增殖和活力的抑制，表明 TCG 可能在 SW480 细胞系中诱导致命的自噬停滞。此外，TCG 在 SW480 细胞系中诱导 YAP 活化。YAP 活性的抑制增强了 TCG 介导的细胞增殖和活力的抑制，表明 YAP 可能在 TCG 诱导的作用中发挥保护作用。总之，本研究的结果表明 TCG 可能诱导致死性自噬停滞并激活 YAP，后者在 SW480 细胞系中起保护作用。这些结果表明，TCG 和 YAP 的联合靶向可能代表了 TCG 介导的抗癌治疗的有前景的策略。