构建胃癌三维多细胞球体模型用于药物瘤内穿透的精准评价  

Constructing Three-Dimensional Multicellular Spheroid Models of Gastric Cancer for Precise Evaluation of Intratumoral Drug Penetration

作　　者：林子鸣 王义俊[1] LIN Ziming;WANG Yijun(Department of Pharmacy,The Second Affiliated Hospital of Nanjing Medical University,Nanjing 210003,China)

机构地区：[1]南京医科大学第二附属医院药学部,南京210003

出　　处：《中国医药指南》2024年第32期1-4,共4页Guide of China Medicine

基　　金：南京医科大学科技发展基金一般项目(NMUB20230005)。

摘　　要：目的构建并表征胃癌三维多细胞球体模型,用于评价化疗药物的瘤内穿透。方法将12000个小鼠胃癌MFC细胞系接种于96 U 3D细胞培养板,以构建MFC多细胞肿瘤球体(MFC MCTS);将8000个MFC细胞和4000个小鼠成纤维NIH/3T3细胞系共同接种于96 U 3D细胞培养板,以构建MFC-NIH/3T3共培养多细胞肿瘤球体(CO MCTS);通过活细胞成像表征球体的直径、圆度、面积和透光率;通过激光共聚焦成像表征球体微环境和化疗药物(LipoDOX,5.0μg/ml)的瘤内穿透。结果培养5 d时,两种球体的透光率最低,圆度>0.90,直径约为530μm,面积约为0.23 mm^(2)。两种球体模型内均检测到3种微环境荧光探针分布。LipoDOX能相对均匀地穿透MFC MCTS,却在CO MCTS内表现出外多内少的不均匀穿透,药物荧光强度降低约58.3%(P<0.01)。结论本研究构建了两种具有良好圆度、高紧实度、适中直径和面积的胃癌三维多细胞球体模型,两种胃癌球体模型均表现出酸性、缺氧和氧化还原微环境,可精准、高效地评价化疗药物的瘤内穿透。Objective To construct and characterize a three-dimensional(3D)multicellular spheroid(MCS)model of gastric cancer to evaluate the penetration of chemotherapeutic drugs within the tumor.Methods A total of 12000 mouse gastric cancer MFC cells were inoculated into a 96 U 3D cell culture plate to construct MFC multicellular tumor spheroids(MFC MCTS);8000 MFC cells and 4000 mouse fibroblast NIH/3T3 cells were co-inoculated into a 96 U 3D cell culture plate to construct MFC-NIH/3T3 co-cultured multicellular tumor spheroids(CO MCTS).The diameter,roundness,area,and transmittance of the spheroids were characterized using live cell imaging.The microenvironment and penetration of the chemotherapeutic drug(LipoDOX,5.0μg/ml)within the spheroids were characterized using laser confocal imaging.Results On day 5 of culture,both types of spheroids had the lowest transmittance,a roundness of>0.90,a diameter of approximately 530μm,and an area of about 0.23 mm^(2).Three types of microenvironmental fluorescence probes were detected within both spheroid models.LipoDOX was able to penetrate relatively evenly through MFC MCTS but showed an uneven penetration in CO MCTS,with the outer part having more drug and the inner part having less,resulting in a reduction of about 58.3%in drug fluorescence intensity(P<0.01).Conclusions This study constructed two types of gastric cancer 3D multicellular spheroid models with good roundness,high compactness,moderate diameter,and area.Both gastric cancer spheroid models exhibited acidic,hypoxic,and redox microenvironments,which can accurately and efficiently evaluate the penetration of chemotherapeutic drugs within the tumor.

关 键 词：胃癌 三维多细胞球体模型 瘤内穿透 化疗药物评价 

分 类 号：R735.2[医药卫生—肿瘤]