声动力联合双重免疫激活作用治疗口腔鳞状细胞癌的研究  

Sonodynamic therapy combined with dual immune activation effect in the treatment of oral squamous cell carcinoma

作　　者：周楠 曹鸣芯 杨晓英[1] ZHOU Nan;CAO Mingxin;YANG Xiaoying(Department of Pharmacy,School of Pharmacy,Tianjin Medical University,Tianjin 300070,China;Department of Orthodontics,Stomatology Hospital,Tianjin Medical University,Tianjin 300070,China)

机构地区：[1]天津医科大学药学院药剂学系,天津300070 [2]天津医科大学口腔医院正畸科,天津300070

出　　处：《天津医科大学学报》2025年第2期167-173,共7页Journal of Tianjin Medical University

基　　金：国家自然科学基金面上项目(12074284)。

摘　　要：目的:探讨声敏剂维替泊芬(Vp)与EZH2抑制剂他泽司他(Taz)构建的纳米药物介导声动力疗法(SDT)与免疫激活协同治疗口腔鳞状细胞癌(OSCC)的作用。方法:通过将Vp与Taz共沉淀制备无载体纳米药物VpTaz,随后通过粒径电位分析仪和透射电镜检测其粒径、粒径分布和形貌特征,并采用活性氧簇(ROS)探针检测其体外SDT性能。进一步采用SCC-7细胞为OSCC细胞的研究对象,观察VpTaz被SCC-7细胞摄取的情况,检测其对胞内ROS生成和细胞毒性的作用,评估其对SCC-7细胞的杀伤性能,通过免疫荧光技术检测SCC-7细胞中高迁移率族蛋白1(HMGB1)的释放情况,考察由VpTaz的SDT作用引发的免疫原性细胞死亡(ICD);同时,通过蛋白印迹实验考察VpTaz对H3K27me3、EZH2、Histone3蛋白表达的影响。结果:VpTaz在透射电子显微镜下呈现完整的实心球形纳米结构,水合粒径大小为(113.03±1.42)nm,分散性良好,多分散系数为0.069±0.016,且具有良好的体外SDT性能(F=1020,P<0.001;F=27.11,P<0.01);其可被SCC-7细胞摄取,并在2 h达到饱和(F=802.1,P<0.001)。在超声激发下,VpTaz处理的SCC-7细胞内产生了大量ROS(F=2550,P<0.001),不同浓度下,非超声Vp、VpTaz组SCC-7细胞其生存率无显著性变化,Vp+US、VpTaz+US组SCC-7细胞的生存率均显著降低(F=5.463、38.53、76.74、398.7、666.7,均P<0.05),且VpTaz可以成功促进HMGB1的释放,促进ICD的发生(F=53.1,P<0.001);在基因层面,对照和单纯Vp相比,Taz和VpTaz均可显著抑制主要组织相容性复合物Ⅰ类分子(MHC-Ⅰ)上游调控相关蛋白的甲基化(F=1006,P<0.01),并且提高MHC-Ⅰ的表达(F=3932,P<0.001),增强SCC-7细胞免疫原性。结论:VpTaz可通过SDT杀伤和双重抗肿瘤免疫激活协同治疗OSCC。Objective:To investigate the synergic effects of nanomedicine mediated sonodynamic therapy(SDT)and antitumor immune activation constructed by Verteporfin(Vp)as a sonosensitizer and Tazemetostat(Taz)as an EZH2 inhibitoron the treatment of oral squa-mous cell carcinoma(OSCC).Methods:Carrier-free nanomedicine VpTaz was obtained by co-precipitation of Vp with Taz,followed by detecting its particle size and size distribution via a particle size potential analyser,and observation the morphological characteristics via transmission electron microscopy.The SDT performance in vitro was detected by reactive oxygen species(ROS)probes.SCC-7 cells were employedas the research object of OSCC cells.The uptake of VpTaz by SCC-7 cells was observed,its effect on intracellular ROS generation and cytotoxicity was detected.Then,the release of high mobility group protein 1(HMGB1)in SCC-7 cells was detected by immunofluorescence technique,the immunogenic death(ICD)triggered by SDT effect of VpTaz was assessed.Meanwhile,the effect of VpTaz on the expression of H3K27me3,EZH2 and Histone3 proteins was examined by Western blotting.Results:VpTaz showed an intact solid spherical nanostructure under transmission electron microscopy,with an average hydrated particle size of(113.03±1.42)nm and well dispersed,a polydispersity coefficient of 0.069±0.016.It exhibited good SDT performance in vitro(F=1020,P<0.001;F=27.11,P<0.01).VpTaz could be uptaken by SCC-7 cells and saturated at 2 h(F=802.1,P<0.001).Under ultrasound irradiation,SCC-7 cells treated with VpTaz exhibited higher ROS levels when compared with OSCC cells without ultrasound irradiation(F=2550,P<0.001).At different concentrations,there was no significant difference in the survival rate of SCC-7 cells in the non ultrasound Vp and VpTaz groups.However,the survival rate of SCC-7 cells in the Vp+US and VpTaz+US groups was significantly reduced(F=5.463,38.53,76.74,398.7,666.7,P<0.05,P<0.001).Moreover,VpTaz could facilitate the release of HMGB1 and induced ICD effect(F=53.1,P<0.001).On the g

关 键 词：口腔鳞状细胞癌 声动力疗法 表观遗传调控 

分 类 号：R739.8[医药卫生—肿瘤]