斑马鱼肝癌细胞与肿瘤相关成纤维细胞可视化转移模型的构建  

Establishment of a visual metastasis model of hepatoma cells and cancer-associated fibroblasts in zebrafish

作　　者：张硕 姜晓燕[1] 沈水杰[2] 吴育[2] 蔡艳[1] 葛炜炜[1] 李高盼 ZHANG Shuo;JIANG Xiaoyan;SHEN Shuijie;WU Yu;CAI Yan;GE Weiwei;LI Gaopan(Department of Pharmacy,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China;Department of Science and Education,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001;Hunter Biotech Company,Hangzhou 310051)

机构地区：[1]南通市中医院药剂科,江苏南通226001 [2]南通市中医院科教科,江苏南通226001 [3]环特生物科技公司,杭州310051

出　　处：《中国实验动物学报》2023年第3期296-301,共6页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(82104408);南通市药学会医院药学基金(ntyx2021)。

摘　　要：目的利用斑马鱼构建肝癌细胞与肿瘤相关成纤维细胞(cancer associated fibroblasts,CAFs)可视化转移模型,为CAFs介导的肿瘤转移患者的机制研究和药效评价提供临床前模型。方法建立肝癌Huh7细胞与CAFs体外共培养体系,分别采用CCK8和Transwell小室检测Huh7细胞的增殖和迁移能力。同时将不同颜色荧光标记的Huh7细胞和CAFs注射到斑马鱼卵黄周隙中,构建可视化转移模型,观察CAFs对Huh7细胞迁移的影响以及Huh7细胞与CAFs在血管中的结合情况。结果与CAFs共培养的Huh7细胞的增殖和迁移能力显著增加。此外,成功建立斑马鱼肝癌细胞和CAFs可视化转移模型。与CAFs共注射的Huh7细胞的迁移能力和结合率显著增加。结论CAFs促进肝癌细胞的迁移,并且在远端转移中,大多数肝癌细胞与CAFs仍处于紧密结合状态。因此在临床上可以靶向实体瘤的成纤维成分,阻断其对肿瘤的支持作用,为肿瘤治疗提供新策略。Objective A visual metastasis model of hepatoma cells and cancer-associated fibroblasts(CAFs)was established using zebrafish to provide a preclinical model for CAFs-mediated mechanism research and therapeutic efficacy evaluation of patients with tumor metastasis.Methods After establishing a co-culture system,the proliferation and migration of Huh7 cells were assessed by CCK8 and Transwell assays,respectively.Huh7 cells and CAFs labeled with different fluorescent dyes were injected into the perivitelline space of zebrafish to establish a visual metastasis model.The effects of CAFs on Huh7 cell migration and Huh7 cell binding to CAFs in blood vessels were observed.Results The proliferation and migration of Huh7 cells co-cultured with CAFs were increased significantly.A visual metastasis model of hepatoma cells and CAFs in zebrafish was established successfully.The migration and binding rate of Huh7 cells co-injected with CAFs were increased significantly.Conclusions CAFs promote migration of hepatoma cells,and most hepatoma cells are tightly bound to CAFs in distal metastases.Therefore,fibroblasts of solid tumors can be targeted to block their support of tumors and provide a new strategy for cancer therapy.

关 键 词：肿瘤相关成纤维细胞 肝细胞肝癌 转移 斑马鱼 可视化 

分 类 号：Q95-33[生物学—动物学]