背景:口腔鳞状细胞癌(OSCC)是最常见的头颈部恶性肿瘤。曾胜平,由山豆根组成。,BistortaofficinalisDelarbre,桑鸡,夏枯草,山药,还有DictamnusdasycarpusTurcz.,被认为是一种具有显著临床疗效的抗癌药物,但因肝脏毒性而停药。本课题组在原增生平的基础上研制出高效低毒的改良增生平(ZSP-M),虽然其药效学机制尚不清楚。这里,我们旨在阐明ZSP-M的药效物质基础,并研究其通过调节肿瘤相关巨噬细胞(TAMs)对OSCC的化学预防作用.
方法:使用超高效液相色谱-质谱法表征ZSP-M的组分。使用4-硝基喹啉N-氧化物癌前病变小鼠模型研究了ZSP-M对实验性口腔癌的化学预防作用。RNA测序分析用于获得ZSP-M处理效果的全局转录视图。细胞共培养模型用于研究ZSP-M对TAM的靶向作用和OSCC细胞的生物学特性,并检测TAM表型的变化。通过分子对接和动态模拟分析了ZSP-M活性化合物与TNFα诱导蛋白6(TNFAIP6)蛋白的结合。
结果:确定了ZSP-M的40种主要成分,其中最丰富的是类黄酮。ZSP-M通过抑制舌癌前病变中TNFAIP6和CD163蛋白的表达来抑制癌前病变中上皮异型增生的程度。ZSP-M抑制增殖,菌落形成,通过靶向TAMs对SCC7细胞的迁移和侵袭。ZSP-M降低CD163+细胞的表达,抑制TNFAIP6蛋白的表达,TAMs中的Arg1mRNA和Il10mRNA,和减少IL-10细胞因子在共培养环境中的释放。在添加重组TNFAIP6蛋白后,这种效果得以维持。计算机模拟表明,三叶根素和马可辛与TNFAIP6连接良好。
结论:ZSP-M通过特异性靶向TNFAIP6来抵消TAM的免疫抑制作用,从而发挥OSCC的化学预防活性。
BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most common malignancy of the head and neck. Zeng-Sheng-Ping, composed of Sophora tonkinensis Gagnep., Bistorta officinalis Delarbre, Sonchus arvensis L., Prunella vulgaris L., Dioscorea bulbifera L., and Dictamnus dasycarpus Turcz., was regarded as an anti-cancer drug with significant clinical efficacy, but was discontinued due to liver toxicity. Our research group developed a modified Zeng-Sheng-Ping (ZSP-M) based on original Zeng-Sheng-Ping that exhibited high efficiency and low toxicity in preliminary investigations, although its pharmacodynamic mechanism is still unclear. Here, we aimed to elucidate the pharmacodynamic material basis of ZSP-M and investigate its chemopreventive effect on OSCC by modulating tumor associated macrophages (TAMs).
METHODS: Components of ZSP-M were characterized using ultra-performance liquid chromatography-mass spectrometry. Chemopreventive effect induced by ZSP-M against experimental oral cancer was investigated using the 4-nitroquinoline N-oxide precancerous lesion mouse model. RNA sequencing analysis was used to gain a global transcriptional view of the effect of ZSP-M treatment. A cell co-culture model was used to study the targeted effect of ZSP-M on TAMs and the biological properties of OSCC cells and to detect changes in TAM phenotypes. The binding of ZSP-M active compounds to TNF alpha induced protein 6 (TNFAIP6) protein was analyzed by molecular docking and dynamic simulation.
RESULTS: Forty main components of ZSP-M were identified, the most abundant of which were flavonoids. ZSP-M inhibited the degree of epithelial dysplasia in precancerous lesions by inhibiting the expression of the TNFAIP6 and CD163 proteins in the precancerous lesions of the tongue. ZSP-M inhibited proliferation, colony formation, migration and invasion of SCC7 cells by targeting TAMs. ZSP-M reduced the expression of CD163+ cells, inhibited the expression of TNFAIP6 protein, Arg1 mRNA and Il10 mRNA in TAMs, and reduced IL-10 cytokine release in the co-culture environment. This effect was maintained after the addition of recombinant TNFAIP6 protein. Computer simulations showed that trifolirhizin and maackiain are well-connected to TNFAIP6.
CONCLUSIONS: ZSP-M counteracts the immunosuppressive action of TAMs by specific targeting of TNFAIP6, thereby exerting chemopreventive activity of OSCC.