进展期胃癌类器官模型的构建和个体化药敏检测  

Construction of advanced gastric cancer organoid model and personalized drug sensitivity testing

作　　者：陈天亮 尹艺霖 彭力达 何梓芸 程菲杨 刘依哲 方效昌 束琳 李炫飞 冯茂辉[1] Chen Tianliang;Yin Yilin;Peng Lida;He Ziyun;Cheng Feiyang;Liu Yizhe;Fang Xiaochang;Su Lin;Li Xuanfei;Feng Maohui(Department of Gastroenterology,Central South Hospital,Wuhan University,Wuhan Clinical Medical Research Center for Peritoneal Cancer,Hubei Provincial Clinical Research Center for Oncology,Hubei Provincial Key Laboratory of Tumor Biological Behavior,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院胃肠外科,武汉市腹膜癌临床医学研究中心,湖北省肿瘤医学临床研究中心,肿瘤生物学行为湖北省重点实验室,武汉430071

出　　处：《中华实验外科杂志》2024年第11期2518-2521,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(82070302、81902018、82472632);武汉市腹膜癌临床医学研究中心资助项目(2015060911020462);吴阶平医学基金会临床科研专项资助项目(320.6750.2023-11-9、320.6750.2023-11-23);武汉大学中南医院科技创新培育基金资助项目(CXPY2022055);武汉大学中南医院医学科技创新平台建设支撑项目资助项目(PTXM2023004、PTXM2023020)。

摘　　要：目的:构建进展期胃癌患者类器官模型,并为其精准化治疗提供一种精准及有效的研究模型。方法:收取10例病理诊断为胃腺癌患者的肿瘤组织标本,通过消化提取肿瘤细胞团,并加入200 μl低生长因子基底膜提取物(BME)平均分配为4~5个液滴进行种板,最后使用特殊胃癌类器官培养基培养7~14 d后进行传代及冻存。通过Image J计数类器官;通过苏木精-伊红(HE)染色比较类器官以及原发组织间组织学特异性;通过药敏实验预测进展期胃癌患者的敏感药物,通过非线性回归(曲线拟合)计算半数抑制浓度(IC 50)。 结果:在收取的10例标本中成功构建出7例进展期胃癌类器官模型,成功率高达70%,其中3例因生长缓慢或无法传代而失败;记录不同模型的类器官数量以及生长状况;HE染色证实类器官模型与原发组织具有相似的组织学特征;药敏试验证明AGCO-1、AGCO-5 、AGCO-8和AGCO-10组对5-氟尿嘧啶(5-Fu)敏感度高于AGCO-2、AGCO-4和AGCO-6组[IC 50分别为(2.753±0.326、3.185±0.165、2.329±0.119)、(3.424±0.155)、(6.438±0.382)、(6.481±0.344)、(5.583±0.394) μmol/L];AGCO-1、AGCO-2、AGCO-8和AGCO-10组对奥沙利铂(Oxaliplatin)敏感度高于AGCO-4、AGCO-5和AGCO-6组[IC 50分别为(5.608±0.270)、(5.338±0.300)、(4.150±0.153)、(5.130±0.021)、(9.303±0.480)、(7.974±0.258)、(7.235±0.333) μmol/L]。 结论:进展期胃癌患者类器官模型构建成功率高达70%,同时类器官模型与原发灶具有一致的组织病理学特征,并可根据药敏实验为患者进行个体化敏感药物治疗的预测与指导。Objective Constructing an organoid model for advanced gastric cancer patients and providing a precise and effective research model for their precision treatment.Methods Collect tumor tissue samples from 10 patients with pathological diagnosis of gastric adenocarcinoma,extract tumor cell clusters through digestion,and add 200μl of low growth factor basement membrane extract(BME)evenly distributed into 4-5 droplets for seeding.Finally,culture in a special gastric cancer organoid culture medium for 7-14 days before passaging and cryopreservation.Counting organoids through image J;Compare the histological specificity between organoids and primary tissues using hematoxylin eosin(HE)staining;Predict the sensitive drugs of advanced gastric cancer patients through drug sensitivity experiments,and calculate the half maximal inhibitory concentration(IC 50)through nonlinear regression(curve fitting).ResultsWe successfully constructed 7 late stage gastric cancer peritoneal metastasis organoid models out of 10 collected specimens,with a success rate of up to 70%.Among them,3 cases failed due to slow growth or inability to passage.We recorded the number and growth status of organoids in different organoid models;HE staining demonstrated that the organoid model has similar histological characteristics to the primary tissue.Drug sensitivity tests have shown that the AGCO-1,AGCO-5,AGCO-8,and AGCO-10 groups are more sensitive to 5-fluorouracil(5-Fu)than the AGCO-2,AGCO-4,and AGCO-6 groups[IC 50 values of(2.753±0.326),(3.185±0.165),(2.329±0.119),(3.424±0.155),(6.438±0.382),(6.481±0.344),(5.583±0.394)μmol/L];The sensitivity of the AGCO-1,AGCO-2,AGCO-8,and AGCO-10 groups to oxaliplatin was higher than that of the AGCO-4,AGCO-5,and AGCO-6 groups[IC 50 values of(5.608±0.270),(5.338±0.300),(4.150±0.153),(5.130±0.021),(9.303±0.480),(7.974±0.258),(7.235±0.333)μmol/L].Conclusion The success rate of constructing organoid models for advanced gastric cancer patients is as high as 70%.At the same time,the organoid model has co

关 键 词：进展期胃癌 类器官模型 精准治疗 

分 类 号：R735.2[医药卫生—肿瘤]