背景:牛黄(CB),用于中药,在各种癌症模型中表现出抗肿瘤作用。它还构成了称为PienTzeHuang的复合制剂的组成部分,用于治疗肝癌。然而,它对肝癌肿瘤微环境的影响,特别是在肿瘤相关巨噬细胞(TAMs)上,不是很了解。
目的:阐明CB通过Wnt/β-catenin通路调控抑制M2-TAM极化的抗肝癌作用。
方法:本研究使用UPLC-Q-TOF-MS鉴定了CB的活性成分,在裸鼠模型中评估了其抗肿瘤作用,并通过网络药理学阐明了潜在的机制,转录组学,和分子对接。体外试验用于研究含CB的血清对HepG2细胞和M2-TAMs的影响。通过实时逆转录酶-聚合酶链反应和Westernblot分析验证了Wnt通路的调节。
结果:这项研究确定了CB中的22种活性成分,其中11在血液中检测到。临床前研究已证明CB有效抑制肝肿瘤生长的能力。采用网络药理学的综合方法,转录组学,和分子对接通过抑制M2-TAM极化将Wnt信号通路作为CB抗肿瘤活性的靶标。体外和体内实验进一步证实,CB显著阻碍M2-TAM极化并抑制Wnt/β-连环蛋白途径活化。当用Wnt激动剂SKL2001处理时,CB对M2-TAM的抑制作用被逆转,证实了其途径特异性。
结论:这项研究表明,CB通过Wnt/β-catenin通路介导M2-TAM极化的抑制,有助于抑制肝癌的生长。
BACKGROUND: Calculus bovis (CB), used in traditional Chinese medicine, exhibits anti-tumor effects in various cancer models. It also constitutes an integral component of a compound formulation known as Pien Tze Huang, which is indicated for the treatment of liver cancer. However, its impact on the liver cancer tumor microenvironment, particularly on tumor-associated macrophages (TAMs), is not well understood.
OBJECTIVE: To elucidate the anti-liver cancer effect of CB by inhibiting M2-TAM polarization via Wnt/β-catenin pathway modulation.
METHODS: This study identified the active components of CB using UPLC-Q-TOF-MS, evaluated its anti-neoplastic effects in a nude mouse model, and elucidated the underlying mechanisms via network pharmacology, transcriptomics, and molecular docking. In vitro assays were used to investigate the effects of CB-containing serum on HepG2 cells and M2-TAMs, and Wnt pathway modulation was validated by real-time reverse transcriptase-polymerase chain reaction and Western blot analysis.
RESULTS: This study identified 22 active components in CB, 11 of which were detected in the bloodstream. Preclinical investigations have demonstrated the ability of CB to effectively inhibit liver tumor growth. An integrated approach employing network pharmacology, transcriptomics, and molecular docking implicated the Wnt signaling pathway as a target of the antineoplastic activity of CB by suppressing M2-TAM polarization. In vitro and in vivo experiments further confirmed that CB significantly hinders M2-TAM polarization and suppresses Wnt/β-catenin pathway activation. The inhibitory effect of CB on M2-TAMs was reversed when treated with the Wnt agonist SKL2001, confirming its pathway specificity.
CONCLUSIONS: This study demonstrated that CB mediates inhibition of M2-TAM polarization through the Wnt/β-catenin pathway, contributing to the suppression of liver cancer growth.