Licorice is a frequently used herbal medicine worldwide, and is used to treat cough, hepatitis, cancer and influenza in clinical practice of traditional Chinese medicine. Modern pharmacological studies indicate that prenylated flavonoids play an important role in the anti-tumor activity of licorice, especially the tumors in stomach, lung, colon and liver. Wighteone is one of the main prenylated flavonoids in licorice, and its possible effect and target against colorectal cancer have not been investigated. This study aimed to investigate the anti-colorectal cancer effect and underlying mechanism of wighteone. SW480 human colorectal cancer cells were used to evaluate the anti-colorectal cancer activity and Akt regulation effect of wighteone by flow cytometry, phosphoproteomic and Western blot analysis. Surface plasmon resonance (SPR) assay, molecular docking and dynamics simulation, and kinase activity assay were used to investigate the direct interaction between wighteone and Akt. A nude mouse xenograft model with SW480 cells was used to verify the anti-colorectal cancer activity of wighteone. Wighteone inhibited phosphorylation of Akt and its downstream kinases in SW480 cells, which led to a reduction in cell viability. Wighteone had direct interaction with both PH and kinase domains of Akt, which locked Akt in a “closed” conformation with allosteric inhibition, and Gln79, Tyr272, Arg273 and Lys297 played the most critical role due to their hydrogen bond and hydrophobic interactions with wighteone. Based on Akt overexpression or activation in SW480 cells, further mechanistic studies suggested that wighteone-induced Akt inhibition led to cycle arrest, apoptosis and autophagic death of SW480 cells. Moreover, wighteone exerted anti-colorectal cancer effect and Akt inhibition activity in the nude mouse xenograft model. Wighteone could inhibit growth of SW480 cells through allosteric inhibition of Akt, which led to cell cycle arrest, apoptosis and autophagic death. The results contributed to understanding of the anti-tumor mechanism of licorice, and also provided a rationale to design novel Akt allosteric inhibitors for the treatment of colorectal cancer.

中文翻译：

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Wighteone 是一种来自甘草的异戊二烯化类黄酮，通过 Akt 的变构抑制来抑制 SW480 结直肠癌细胞的生长

甘草是世界范围内常用的草药，在中医临床中用于治疗咳嗽、肝炎、癌症和流感。现代药理研究表明，异戊烯化黄酮类化合物在甘草的抗肿瘤活性中发挥着重要作用，特别是对胃、肺、结肠和肝等肿瘤的抑制作用。 Wighteone是甘草中主要的异戊二烯化黄酮类化合物之一，其对抗结直肠癌的可能作用和靶点尚未得到研究。本研究旨在探讨wighteone的抗结直肠癌作用及其潜在机制。采用SW480人结直肠癌细胞，通过流式细胞术、磷酸化蛋白质组学和Western blot分析评价wighteone的抗结直肠癌活性和Akt调节作用。采用表面等离子共振 (SPR) 测定、分子对接和动力学模拟以及激酶活性测定来研究 wighteone 和 Akt 之间的直接相互作用。采用SW480细胞裸鼠异种移植模型验证wighteone的抗结直肠癌活性。 Wighteone 抑制 SW480 细胞中 Akt 及其下游激酶的磷酸化，从而导致细胞活力降低。 Wighteone 与 Akt 的 PH 和激酶结构域均具有直接相互作用，从而将 Akt 锁定在“封闭”构象并具有变构抑制作用，而 Gln79、Tyr272、Arg273 和 Lys297 由于其氢键和与 Wighteone 的疏水相互作用而发挥着最关键的作用。基于 SW480 细胞中 Akt 的过度表达或激活，进一步的机制研究表明 wighteone 诱导的 Akt 抑制导致 SW480 细胞的周期停滞、细胞凋亡和自噬性死亡。此外，wighteone 在裸鼠异种移植模型中发挥抗结直肠癌作用和 Akt 抑制活性。 Wighteone 可以通过 Akt 的变构抑制来抑制 SW480 细胞的生长，从而导致细胞周期停滞、细胞凋亡和自噬死亡。该结果有助于了解甘草的抗肿瘤机制，也为设计用于治疗结直肠癌的新型 Akt 变构抑制剂提供了理论基础。