白藜芦醇调控乳酸代谢重编程介导的肿瘤相关巨噬细胞极化在喉癌中的作用机制

Mechanism of Action of Resveratrol in Lactate Metabolism Reprogramming-Mediated Tumor-Associated Macrophage Polarization in Laryngeal Cancer

摘要:

目的：探讨白藜芦醇(resveratrol, RES)是否通过调控乳酸代谢重编程介导的肿瘤相关巨噬细胞(tumor-associated macrophages, TAMs)极化在喉癌中发挥抗肿瘤作用,并阐明其潜在分子机制。方法：采用Hep-2细胞与THP-1细胞共培养体系模拟肿瘤免疫微环境。通过Western blot检测MCT1表达及ERK1/2、STAT3信号通路磷酸化水平；ELISA检测TNF-α、IL-10和TGF-β水平；免疫荧光检测CD86和CD206表达。构建Hep-2皮下移植瘤裸鼠模型,观察肿瘤生长、巨噬细胞极化状态及组织乳酸水平变化。结果：与Hep-2细胞共培养后,巨噬细胞MCT1表达、p-ERK1/2及p-STAT3水平显著升高,TNF-α减少而IL-10和TGF-β增加, CD206表达上调,Hep-2细胞活力增强。RES干预逆转上述变化,显著下调MCT1表达,抑制ERK1/2和STAT3磷酸化,促进M1型极化并抑制M2型极化,降低喉癌细胞活力。动物实验表明,RES显著抑制共接种组肿瘤生长,降低瘤内乳酸水平和M2型巨噬细胞比例。乳酸钠干预可部分抵消RES对ERK/STAT3信号及肿瘤生长的抑制作用。结论：喉癌细胞来源的乳酸通过MCT1作用于巨噬细胞激活ERK/STAT3通路,诱导其向M2型极化。RES通过下调巨噬细胞MCT1表达,阻断乳酸代谢通讯,抑制ERK/STAT3信号通路激活,逆转M2型极化,从而抑制喉癌进展。

Abstract:

Objective: To investigate whether resveratrol (RES) exerts an anti-tumor effect in laryngeal cancer by regulating lactate metabolism reprogramming-mediated tumor-associated macrophage (TAM) polarization, and to elucidate its potential molecular mechanism. Methods: A co-culture system of Hep-2 cells and THP-1 cells was used to simulate the tumor immune microenvironment. Western blot was used to detect MCT1 expression and phosphorylation levels of ERK1/2 and STAT3 signaling pathways; ELISA was used to detect TNF-α, IL-10, and TGF-β levels; and immunofluorescence was used to detect CD86 and CD206 expression. A Hep-2 subcutaneous xenograft tumor model was established in nude mice to observe tumor growth, macrophage polarization status, and changes in tissue lactate levels. Results: Co-culturing macrophages with Hep-2 cells significantly increased MCT1 expression, p-ERK1/2 and p-STAT3 levels, decreased TNF-α while increasing IL-10 and TGF-β, upregulated CD206 expression, and enhanced Hep-2 cell viability. RES intervention reversed these changes, significantly downregulating MCT1 expression, inhibiting ERK1/2 and STAT3 phosphorylation, promoting M1 polarization and inhibiting M2 polarization, and reducing laryngeal cancer cell viability. Animal experiments showed that RES significantly inhibited tumor growth in the co-inoculation group, reduced intratumoral lactate levels and the proportion of M2 macrophages. Sodium lactate intervention partially counteracted the inhibitory effects of RES on ERK/STAT3 signaling and tumor growth. Conclusion: Lactate derived from laryngeal cancer cells activates the ERK/STAT3 pathway in macrophages via MCT1, inducing M2 polarization. RES inhibits this process by downregulating MCT1 expression in macrophages, thereby blocking lactate-mediated metabolic communication and suppressing laryngeal cancer progression.