背景:StephaniacephananthaHayata是一种重要的传统药用植物,在传统医学中广泛用于治疗癌症。从StephaniacephalanthaHayata的根中提取了头孢三氨酸(CEP)。已发现其在不同类型的癌细胞中表现出抗癌活性。然而,CEP对鼻咽癌(NPC)的活性及其潜在机制有待进一步研究。
目的:NPC是一种侵袭性高转移性恶性肿瘤,影响头颈部。本研究旨在探讨CEP抗鼻咽癌的药理特性和潜在机制。旨在为使用CEP治疗NPC提供新的观点。
方法:体外,使用CCK-8试验评估CEP对NPC的药理活性。预测和阐明CEP对鼻咽癌的抗癌机制,我们采用了网络药理学,进行了分子对接分析,并进行蛋白质印迹实验。通过人NPC的裸小鼠异种移植模型进行体内验证,蛋白质印迹和免疫组织化学(IHC)测定以确认药理活性和机制。
结果:以剂量依赖性方式,CEP显著抑制NPC细胞的增殖和克隆形成能力。此外,NPC细胞迁移被CEP抑制。网络药理学实验结果表明,CEP的抗NPC效应与8个核心靶点有关,包括EGFR,AKT1,PIK3CA,还有MTOR.通过进行分子对接,CEP与候选核心蛋白(EGFR,AKT1,PIK3CA,和mTOR)被预测,导致EGFR的对接能量为-10.0kcal/mol,-12.4kcal/mol的PIK3CA,AKT1为-10.8kcal/mol,mTOR为-8.6kcal/mol。Westernblot分析显示CEP能有效抑制EGFR的表达和下游信号蛋白的磷酸化水平,包括PI3K,AKT,mTOR,和ERK.CEP干预后,肿瘤大小的显著减小,不引起任何毒性,在经历体内治疗的NPC异种移植裸鼠中观察到。此外,IHC分析证实CEP治疗后EGFR和Ki-67的表达水平显著降低。
结论:CEP对鼻咽癌具有显著的药理作用,其机制作用涉及抑制EGFR/PI3K/AKT通路的激活。CEP代表用于解决和减轻NPC的有前途的药剂。
BACKGROUND: The Stephania cephalantha Hayata is an important traditional medicinal plant widely used in traditional medicine to treat cancer.
Cepharanthine (CEP) was extracted from the roots of Stephania cephalantha Hayata. It has been found to exhibit anticancer activity in different types of cancer cells. Nevertheless, the activity of CEP against nasopharyngeal carcinoma (NPC) and its underlying mechanism warrant further investigation.
OBJECTIVE: NPC is an invasive and highly metastatic malignancy that affects the head and neck region. This research aimed to investigate the pharmacological properties and underlying mechanism of CEP against NPC, aiming to offer novel perspectives on treating NPC using CEP.
METHODS: In vitro, the pharmacological activity of CEP against NPC was evaluated using the CCK-8 assay. To predict and elucidate the anticancer mechanism of CEP against NPC, we employed network pharmacology, conducted molecular docking analysis, and performed Western blot experiments. In vivo validation was performed through a nude mice xenograft model of human NPC, Western blot and immunohistochemical (IHC) assays to confirm pharmacological activity and the mechanism.
RESULTS: In a dose-dependent manner, the proliferation and clonogenic capacity of NPC cells were significantly inhibited by CEP. Additionally, NPC cell migration was suppressed by CEP. The results obtained from network pharmacology experiments revealed that anti-NPC effect of CEP was associated with 8 core targets, including EGFR, AKT1, PIK3CA, and mTOR. By performing molecular docking, the binding capacity of CEP to the candidate core proteins (EGFR, AKT1, PIK3CA, and mTOR) was predicted, resulting in docking energies of -10.0 kcal/mol for EGFR, -12.4 kcal/mol for PIK3CA, -10.8 kcal/mol for AKT1, and -8.6 kcal/mol for mTOR. The Western blot analysis showed that CEP effectively suppressed the expression of EGFR and the phosphorylation levels of downstream signaling proteins, including PI3K, AKT, mTOR, and ERK. After CEP intervention, a noteworthy decrease in tumor size, without inducing any toxicity, was observed in NPC xenograft nude mice undergoing in vivo treatment. Additionally, IHC analysis demonstrated a significant reduction in the expression levels of EGFR and Ki-67 following CEP treatment.
CONCLUSIONS: CEP exhibits significant pharmacological effects on NPC, and its mechanistic action involves restraining the activation of the EGFR/PI3K/AKT pathway. CEP represents a promising pharmaceutical agent for addressing and mitigating NPC.