中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (34): 8970-8977.doi: 10.12307/2026.891

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

患者源性食管胃结合部腺癌类器官模型的构建与鉴定

高震东1，2，张  理2，陈  攀2，尹希瑶2，李佳怡2，杨平娟2，张  敏1，廖淑馨2，石林林2，高社干2   

  1. 1河南科技大学基础医学与法医学院，河南省洛阳市  471023；2河南科技大学临床医学院，河南科技大学第一附属医院，河南省微生态与食管癌防治重点实验室，河南省洛阳市  471003
  • 收稿日期:2025-09-23 修回日期:2026-02-14 出版日期:2026-12-08 发布日期:2026-04-14
  • 通讯作者: 石林林，博士，副教授，副主任医师，河南科技大学临床医学院，河南科技大学第一附属医院，河南省微生态与食管癌防治重点实验室，河南省洛阳市 471003 共同通讯作者：高社干，博士，教授，主任医师，河南科技大学临床医学院，河南科技大学第一附属医院，河南省微生态与食管癌防治重点实验室，河南省洛阳市 471003
  • 作者简介:高震东，男，2000年生，河南省新乡市人，汉族，硕士，主要从事消化道肿瘤临床与基础研究。
  • 基金资助:
    国家自然科学基金项目(82302966)，项目负责人：石林林；国家临床重点专科建设项目联合基金(ZLKFJJ20230401)，项目负责人：石林林；河南省临床医学科学家培养专项(HNCMS202438)，项目负责人：石林林；河南省医学科技攻关省部共建重点项目(LHGJ20230455)，项目负责人：陈攀

Establishment and identification of a patient-derived organoid model for esophagogastric junction adenocarcinoma

Gao Zhendong1, 2, Zhang Li2, Chen Pan2, Yin Xiyao2, Li Jiayi2, Yang Pingjuan2, Zhang Min1, Liao Shuxin2, Shi Linlin2, Gao Shegan2   

  1. 1School of Basic Medical Sciences and Forensic Medicine, Henan University of Science and Technology, Luoyang 471023, Henan Province, China; 2The First Affiliated Hospital, College of Clinical Medicine, Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment, Henan University of Science and Technology, Luoyang 471003, Henan Province, China
  • Received:2025-09-23 Revised:2026-02-14 Online:2026-12-08 Published:2026-04-14
  • Contact: Shi Linlin, PhD, Associate professor, Associate chief physician, The First Affiliated Hospital, College of Clinical Medicine, Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment, Henan University of Science and Technology, Luoyang 471003, Henan Province, China Co-corresponding author: Gao Shegan, PhD, Professor, Chief physician, The First Affiliated Hospital, College of Clinical Medicine, Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment, Henan University of Science and Technology, Luoyang 471003, Henan Province, China
  • About author:Gao Zhendong, MS, School of Basic Medical Sciences and Forensic Medicine, Henan University of Science and Technology, Luoyang 471023, Henan Province, China; The First Affiliated Hospital, College of Clinical Medicine, Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment, Henan University of Science and Technology, Luoyang 471003, Henan Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 82302966 (to SLL); Joint Fund of National Key Clinical Specialty Construction Project, No. ZLKFJJ20230401 (to SLL); Henan Provincial Training Program for Clinical Medical Scientists, No. HNCMS202438 (to SLL); Key Project of Henan Provincial Medical Science and Technology Research, No. LHGJ20230455 (to CP)

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摘要：


文题释义：
食管胃结合部腺癌：是起源于食管与胃交界区域(通常指食管下括约肌上下5 cm范围)的恶性肿瘤，具有隐匿性强、异质性高等特点，多数患者确诊时病情已进展至中晚期，治疗响应差且总体生存率较低，是目前消化道肿瘤基础研究与临床诊疗工作的关键难点问题。 
类器官：是由成体干细胞或多能干细胞在体外三维培养体系中增殖分化形成的微型组织类似物，具有与来源组织高度相似的结构特征、细胞组成及功能特性，可模拟体内组织的生理病理状态，为疾病建模、药物筛选、机制研究及个体化医疗提供理想的体外模型工具。 

背景：传统细胞培养模型在精准模拟肿瘤生物学特性方面存在较大局限，类器官技术凭借保留原发肿瘤关键生物学特性的优势，为深入探究食管胃结合部腺癌的发病机制以及开展精准诊疗相关研发工作提供技术支撑。
目的：构建患者源性食管胃结合部腺癌类器官模型，评估与原发肿瘤生物学特征的一致性，为食管胃结合部腺癌的发病机制研究及精准诊疗提供标准化体外模型。
方法：收集50例食管胃结合部腺癌患者手术切除标本进行组织分离、培养及类器官模型构建；采用荧光共定位染色分析类器官模型中的细胞活性；通过苏木精-伊红染色分析类器官模型与原发肿瘤组织的结构特征；运用过碘酸雪夫染色检测类器官模型中的黏液分泌表型；采用免疫组织化学染色评估类器官模型与原发肿瘤组织病理学的一致性；使用卡方检验及单因素逻辑回归分析类器官模型构建成功率与患者临床病理特征、组织特征的相关性。
结果与结论：成功构建了患者源性食管胃结合部腺癌类器官模型培养体系，原代培养成功率为86.0%(43/50)，类器官呈典型三维结构，自细胞团块逐步发育为腺腔样三维实性球体，经原代培养7-10 d后可稳定传代，连续传代多次未出现明显表型改变，且冻存复苏后类器官仍可稳定增殖；荧光共定位染色显示类器官模型在不同培养阶段均可保持较高活力；苏木精-伊红染色结果显示类器官模型与原发肿瘤组织均呈现异型腺体排列及高核质比特征；过碘酸雪夫染色同步呈现阳性黏液分泌；免疫组织化学染色显示类器官模型与原发肿瘤组织的Ki67、CEA、CK7、Cadherin17阳性表达率具有高度一致性。此外，卡方检验及逻辑回归分析结果显示，肿瘤组织低分化(P=0.02)、经新辅助化疗达到主要病理缓解(P=0.007)、肿瘤组织离体时间(P=0.006)及肿瘤质量(P=0.006)可显著影响患者源性食管胃结合部腺癌类器官模型构建的成功率。

https://orcid.org/0000-0001-6903-0281(石林林)；https://orcid.org/0000-0001-7891-9349(高社干)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 食管胃结合部腺癌, 原代培养, 三维培养, 类器官, 模型构建, 细胞活力, 组织病理分析, 肿瘤模型

Abstract: BACKGROUND: Traditional cell culture models exhibit substantial limitations in accurately recapitulating the complex biological characteristics of tumors. In contrast, organoid, which maintains the essential biological features of primary tumors, offers a highly effective platform for comprehensive investigation into the pathogenesis of adenocarcinoma of the esophagogastric junction and the advancement of precision diagnostics and targeted therapeutic strategies.
OBJECTIVE: To Establish patient-derived organoid models of gastroesophageal junction adenocarcinoma, evaluate their fidelity in representing the biological characteristics of primary tumors, and provide a standardized in vitro model for investigating the pathogenesis of gastroesophageal junction adenocarcinoma and advancing precision medicine.
METHODS: Fifty surgical resection specimens were obtained from patients with adenocarcinoma of the esophagogastric junction and used for tissue dissociation, culture, and the establishment of organoid models. Cell viability in the constructed organoid models was evaluated using fluorescence co-localization staining. Hematoxylin-eosin staining was performed to compare structural morphological features between patient-derived organoid models of esophagogastric junction adenocarcinoma and their corresponding primary tumor tissues. Periodic Acid-Schiff staining was carried out to detect mucin-secreting phenotypes in the patient-derived organoid models. Immunohistochemistry was conducted to assess histopathological consistency between the organoid models and the original tumor tissues. The chi-square test and univariate logistic regression analysis were utilized to explore potential associations between the success rate of organoid model establishment and the clinicopathological characteristics as well as tissue features of the patients.
RESULTS AND CONCLUSION: In the patient-derived organoid models of esophagogastric junction adenocarcinoma established in this study, the primary culture success rate was 86.0% (43/50). The organoids exhibited a typical three-dimensional structure, progressively developing from cell aggregates into glandular lumen-like spherical formations. Stable subculturing was achieved within 7-10 days after primary culture, with no significant phenotypic changes observed across multiple passages. Furthermore, the organoids maintained a stable proliferative capacity after cryopreservation and subsequent thawing. Fluorescence co-localization staining confirmed that the patient-derived organoid models of esophagogastric junction adenocarcinoma maintained high cell viability across all stages of culture. Hematoxylin-eosin staining revealed that both the patient-derived organoid models and the corresponding primary tumor tissues exhibited dysplastic glandular architecture and an increased nuclear-to-cytoplasmic ratio. Periodic Acid-Schiff staining demonstrated consistent mucin secretion positivity. Immunohistochemistry analysis revealed a high degree of concordance in the positive expression rates of Ki67, CEA, CK7, and Cadherin17 between the organoids and the corresponding primary tumor tissues. Moreover, chi-square test and logistic regression analyses indicated that poorly differentiated tumor tissues (P=0.02), major pathological response to neoadjuvant chemotherapy in patients with adenocarcinoma of the esophagogastric junction (P=0.007), ex vivo duration of tumor tissues (P=0.006), and tumor tissue weight (P=0.006) were significantly associated with the success rate of organoid model establishment.


Key words: adenocarcinoma of the esophagogastric junction, primary culture, three-dimensional culture, organoids, model establishment, cell viability, histopathological analysis, tumor model

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