5-FU节拍化疗策略优化及调节胃癌免疫微环境的体内实验研究  

Optimization of 5-FU metronomic chemotherapy strategy and regulation of the immune microenvironment in gastric cancer:an in vivo study

作　　者：蒋金玲[1] 周尘飞[1,2] 奚文崎[1] 施敏[1] 耿梅[1] 赵丽琴 蔡劬[1] 蒋劲松[1] 张俊[1,2] JIANG Jinling;ZHOU Chenfei;XI Wenqi;SHI Min;GEN Mei;ZHAO Liqin;CAI Qu;JIANG Jinsong;ZHANG Jun(Department of Oncology,Ruijin Hospital,Shanghai Jiao Tong University,Shanghai 200025,China;Department of Oncology,Wuxi Branch of Ruijin Hospital Affiliated to Medical College of Shanghai Jiao Tong University,Wuxi 214021,Jiangsu Province,China)

机构地区：[1]上海交通大学医学院附属瑞金医院肿瘤科,上海200025 [2]上海交通大学医学院附属瑞金医院无锡分院肿瘤科,江苏无锡214021

出　　处：《中国癌症杂志》2022年第7期596-605,共10页China Oncology

基　　金：国家自然科学基金(81972707,81502013);上海市卫生健康委协同创新集群计划(2020CXJQ03)。

摘　　要：背景与目的:5-氟尿嘧啶(5-fluorouracil,5-FU)是胃癌化疗的骨架药物,传统大剂量5-FU常导致严重不良反应及耐药。低剂量5-FU节拍化疗在不影响疗效的前提下可明显降低药物毒性,但何种给药节拍可达到最佳抗肿瘤作用尚不清楚。本研究旨在探索5-FU的最佳节拍化疗模式,并研究其对胃癌免疫微环境的影响。方法:建立SGC-7901胃癌细胞系的BALB/c裸小鼠皮下移植瘤模型,成瘤后随机分成4组:最大耐受剂量(maximum tolerated dose,MTD)组、每日节拍化疗(daily metronomic chemotherapy,MET-qd)组、隔日节拍化疗(every other day metronomic chemotherapy,MET-qod)组和每周2次节拍化疗(twice-weekly metronomic chemotherapy,MET-biw)组。21 d为1个疗程,共2个疗程。治疗期间观测裸小鼠的一般状况,每周称重并测量瘤体,绘制肿瘤生长曲线。采用流式细胞术检测裸小鼠外周血内皮祖细胞(circulating endothelial progenitors,CEP),瘤体及脾脏内浸润的B细胞、自然杀伤(natural killer,NK)细胞、肿瘤相关巨噬细胞(tumor-associated macrophage,TAM)、髓源性抑制细胞(myeloid-derived suppressor cell,MDSC)。采用免疫组织化学染色法检测瘤体内CD11c和CD163蛋白的表达。采用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测裸小鼠外周血中血管内皮生长因子(vascular endothelial growth factor,VEGF)、血小板源性生长因子(platelet derived growth factor,PDGF)、白细胞介素(interleukin,IL)-10和IL-12的表达。采用白细胞计数及H-E染色等评价肝、肺、肾和心脏毒性。结果:5-FU的3种节拍化疗模式均显示出与MTD组类似的抑制裸小鼠移植瘤生长效应,其中MET-qod组的抗肿瘤效应最明显(P<0.05)。与MTD组(45.3%±4.3%)相比,5-FU的3种节拍化疗模式均可明显降低裸小鼠外周血的CEP数量,其中MET-qd组降低最明显(14.8%±3.8%)。外周血中VEGF在MET-qod组中下降最明显(P<0.001),而在MET-biw组中则明显升高(P<0.001)。PDGF的表�Background and purpose:5-fluorouracil(5-FU)is the backbone of drug for gastric cancer.Traditional high-dose 5-FU often leads to serious adverse reactions and drug resistance.The low-dose 5-FU metronomic chemotherapy can significantly reduce the toxicity of the drug without affecting the efficacy,but it is not clear which dose can achieve the best anti-tumor effect.The purpose of this study was to explore the optimal metronomic chemotherapy strategy for 5-FU and to explore the related molecular mechanisms.Methods:A nude mouse subcutaneous tumor model of SGC-7901 gastric cancer cell line was established.After tumor formation,mice were randomly divided into 4 groups:maximum tolerated dose(MTD)group,daily metronomic chemotherapy(MET-qd)group,every other day metronomic chemotherapy(MET-qod)group and twice-weekly metronomic chemotherapy(MET-biw)group.Duration of single course of treatment was 21 d,and mice received a total of 2 courses of treatment.During the treatment period,the general condition of nude mice was observed,the tumor mass was weighed and measured every week,and the tumor growth curve was drawn.Flow cytometry was used to detect circulating endothelial progenitors(CEP)in peripheral blood of nude mice;B cells,natural killer(NK)cells,tumor-associated macrophage(TAM)and myeloid-derived suppressor cell(MDSC)infiltrated in the tumor and spleen.Immunohistochemical staining was used to detect the expressions of CD11c and CD163 in the tumor.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of vascular endothelial growth factor(VEGF),platelet derived growth factor(PDGF),interleukin(IL)-10 and IL-12 in peripheral blood of nude mice.In addition,white blood cell counts and H-E stained sections of liver,lung,kidney,and heart were performed in order to monitor toxicity assessments.Results:The three metronomic chemotherapy strategies of 5-FU could inhibit the growth of xenograft tumor in nude mice similar to the MTD group,and the MET-qod group has the most obvious anti-tumor effect(P<0.05).Compa

关 键 词：5-氟尿嘧啶 节拍化疗 胃癌 肿瘤相关巨噬细胞 抗血管生成 

分 类 号：R735.2[医药卫生—肿瘤]