背景:根据中医气血理论,补气活血中药合用对改善血瘀证具有协同作用,尤其是在肿瘤治疗中。经典的“黄芪-丹参”二重奏体现了这一原则,以活气和活血而闻名,广泛应用于肿瘤治疗。我们先前的研究强调了Formononetin(来自黄芪)和丹酚酸B(来自丹参)在体外的组合对胰腺肿瘤异种移植物的有效抑制作用。然而,目前尚不清楚这种组合是否可以抑制胰腺肿瘤的异常血管化,从而达到其抗癌作用。
目标:血管异常,已知促进肿瘤生长和转移。使肿瘤相关血管正常化的策略为抗肿瘤治疗提供了有希望的途径。本研究旨在揭示Formononetin联合丹酚酸B(FcS)对胰腺癌内皮细胞影响的治疗潜力。阐明了管理这种治疗相互作用的潜在机制,从而推进使肿瘤脉管系统正常化和对抗癌症进展的策略。
方法:建立了涉及人脐静脉内皮细胞(HUVECs)和PANC-1细胞的共培养系统,以研究靶向异常脉管系统作为新型抗肿瘤治疗策略的潜力。我们系统地比较了HUVEC的增殖,迁移,入侵,在与PANC-1(H-P)的单培养和共培养条件下的腔内发生。随后,评估H-P系统的FcS治疗的抗血管生成性质。利用分子对接预测了Formononetin和丹酚酸B与RhoA的相互作用,并评估了治疗后HUVECs中RhoA的表达。此外,我们利用shRhoA慢病毒阐明了RhoA在FcS介导的HUVECs效应中的作用。在体内,斑马鱼异种移植肿瘤模型用于评估FcS的抗肿瘤潜力,专注于癌细胞增殖,迁移,凋亡,和血管发育。
结果:FcS治疗表明,剂量依赖性抑制PANC-1诱导的HUVECs改变,包括扩散,迁移,入侵,和管道形成能力。分子对接分析表明FcS和RhoA之间的潜在相互作用。Further,发现FcS治疗下调PANC-1诱导的HUVEC中的RhoA表达并调节PI3K/AKT信号通路。值得注意的是,FcS对HUVECs的表型抑制作用通过RhoA敲低减弱。体内斑马鱼研究验证了FcS的抗肿瘤活性,抑制癌细胞增殖,转移,和血管发芽,同时促进肿瘤细胞凋亡。
结论:本研究强调了FcS在对抗胰腺癌诱导的内皮改变方面的潜在潜力。FcS表现出明显的抗异常血管效应,可能通过下调RhoA和抑制PI3K/Akt信号通路来实现,从而为胰腺癌的治疗提供了新的治疗途径。
BACKGROUND: According to the theory of Qi and blood in Traditional Chinese Medicine (TCM), the combination of Qi-reinforcing herbs and blood-activating herbs has a synergistic effect in improving blood stasis syndrome, especially in tumor treatment. The classic \"Radix Astragali - Salvia miltiorrhiza\" duo exemplifies this principle, renowned for invigorating Qi and activating blood flow, employed widely in tumor therapies. Our prior research underscores the potent inhibition of pancreatic tumor xenografts by the combination of Formononetin (from Radix Astragali) and Salvianolic acid B (from Salvia miltiorrhiza) in vitro. However, it remains unclear whether this combination can inhibit the abnormal vascularization of pancreatic tumors to achieve its anti-cancer effect.
OBJECTIVE: Abnormal vasculature, known to facilitate tumor growth and metastasis. Strategies to normalize tumor-associated blood vessels provide a promising avenue for anti-tumor therapy. This study aimed to unravel the therapeutic potential of Formononetin combined with Salvianolic acid B (FcS) in modulating pancreatic cancer\'s impact on endothelial cells, illuminate the underlying mechanisms that govern this therapeutic interaction, thereby advancing strategies to normalize tumor vasculature and combat cancer progression.
METHODS: A co-culture system involving Human Umbilical Vein Endothelial Cells (HUVECs) and PANC-1 cells was established to investigate the potential of targeting abnormal vasculature as a novel anti-tumor therapeutic strategy. We systematically compared HUVEC proliferation, migration, invasion, and lumenogenesis in both mono- and co-culture conditions with PANC-1 (H-P). Subsequently, FcS treatment of the H-P system was evaluated for its anti-angiogenic properties. Molecular docking was utilized to predict the interactions between Formononetin and Salvianolic acid B with RhoA, and the post-treatment expression of
RhoA in HUVECs was assessed. Furthermore, we utilized shRhoA lentivirus to elucidate the role of
RhoA in FcS-mediated effects on HUVECs. In vivo, a zebrafish xenograft tumor model was employed to assess FcS\'s anti-tumor potential, focusing on cancer cell proliferation, migration, apoptosis, and vascular development.
RESULTS: FcS treatment demonstrated a significant, dose-dependent inhibition of PANC-1-induced alterations in HUVECs, including proliferation, migration, invasion, and tube formation capabilities. Molecular docking analyses indicated potential interactions between FcS and
RhoA. Further, FcS treatment was found to downregulate
RhoA expression and modulated the PI3K/AKT signaling pathway in PANC-1-induced HUVECs. Notably, the phenotypic inhibitory effects of FcS on HUVECs were attenuated by
RhoA knockdown. In vivo zebrafish studies validated FcS\'s anti-tumor activity, inhibiting cancer cell proliferation, metastasis, and vascular sprouting, while promoting tumor cell apoptosis.
CONCLUSIONS: This study underscores the promising potential of FcS in countering pancreatic cancer-induced endothelial alterations. FcS exhibits pronounced anti-abnormal vasculature effects, potentially achieved through downregulation of RhoA and inhibition of the PI3K/Akt signaling pathway, thereby presenting a novel therapeutic avenue for pancreatic cancer management.