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7～14d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(IC50).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[IC50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[IC50 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.
Abstract
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 tis-sue samples from 10 patients with pathological diagnosis of gastric adenocarcinoma,extract tumor cell clus-ters through digestion,and add 200 µ l of low growth factor basement membrane extract(BME)evenly dis-tributed into 4-5 droplets for seeding.Finally,culture in a special gastric cancer organoid culture medium for 7-14 days before passaging and cry opreservation.Counting organoids through image J;Compare the his-tological 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(IC50)through nonlinear regression(curve fitting).Results We 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[IC50 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[IC50 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 consistent histopatho-logical characteristics with the primary lesion and can predict and guide personalized sensitive drug treat-ment for patients based on drug sensitivity experiments.
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