Cancer-associated fibroblasts promote proliferation and drug resistance of gastric cancer organoids: a primary study

ZHANG Yuanyuan; DUAN Zhenquan; LI Yuxian; HUANG Mengqiu; ZHU Baohang; Qiu Yuan; ZOU Quanming; PENG Liusheng; MA Daiyuan

doi:10.16016/j.2097-0927.202410075

您当前的位置：

首页 >

文章列表页 >

Cancer-associated fibroblasts promote proliferation and drug resistance of gastric cancer organoids: a primary study

Monographic Reports | 更新时间：2025-03-11

- Cancer-associated fibroblasts promote proliferation and drug resistance of gastric cancer organoids: a primary study
- Journal of Army Medical University Vol. 47, Issue 5, Pages: 453-461(2025)
- 作者机构：
  
  1.川北医学院附属医院肿瘤科，四川南充
  2.陆军军医大学（第三军医大学）药学与检验医学系微生物与生化药学教研室，重庆
  3.陆军军医大学（第三军医大学）第二附属医院普外科，重庆
- 作者简介：
- 基金信息：
  
  the Beijing Science and Technology Innovation Medical Development Foundation(KC2023-JX-0186-FM057);the Scientific Research Development Program of the Affiliated Hospital of North Sichuan Medical College(2023PTZK014)
- DOI：10.16016/j.2097-0927.202410075
  CLC： R329.24;R73-354;R735.2
- Received：22 October 2024，
  
  Revised：27 January 2025，
  
  Published：15 March 2025
- 稿件说明：
移动端阅览
张媛媛，段振铨，李雨贤，等.肿瘤相关成纤维细胞促胃癌类器官增殖与耐药的初步研究［J］.陆军军医大学学报，2025，47（05）：453-461.DOI：10.16016/j.2097-0927.202410075.

ZHANG Y Y，DUAN Z Q，LI Y X，et al.Cancer-associated fibroblasts promote proliferation and drug resistance of gastric cancer organoids： a primary study［J］.J Army Med Univ，2025，47（05）：453-461.DOI：10.16016/j.2097-0927.202410075.
张媛媛，段振铨，李雨贤，等.肿瘤相关成纤维细胞促胃癌类器官增殖与耐药的初步研究［J］.陆军军医大学学报，2025，47（05）：453-461.DOI：10.16016/j.2097-0927.202410075. DOI：

ZHANG Y Y，DUAN Z Q，LI Y X，et al.Cancer-associated fibroblasts promote proliferation and drug resistance of gastric cancer organoids： a primary study［J］.J Army Med Univ，2025，47（05）：453-461.DOI：10.16016/j.2097-0927.202410075. DOI：

摘要

目的

体外构建胃癌类器官与肿瘤相关成纤维细胞的共培养模型，利用该模型探究肿瘤相关成纤维细胞对胃癌类器官增殖与化疗耐药的影响。

方法

收集于2023年2月至2024年3月在陆军军医大学第二附属医院普外科接受手术切除的12例胃癌患者的肿瘤组织。采用体外三维构建胃癌类器官；通过HE染色进行形态学分析，采用免疫组织化学染色检测细胞角蛋白CK7、癌胚抗原CEA和增殖相关因子Ki-67的表达。培养同一患者来源的肿瘤相关成纤维细胞，通过光镜观察分析其形态；采用流式细胞染色检测其表型。将胃癌类器官与肿瘤相关成纤维细胞按1∶1比例共培养，相差显微镜下观察胃癌类器官的生长情况并分析其数量、平均直径与总面积。将单独培养的类器官设置为对照组，在对照组和共培养组中加入化疗药物5-氟尿嘧啶和奥沙利铂处理48 h，利用CellTiter-Glo

3D试剂盒和CellEvent™ Caspase 3/7 分别检测类器官的细胞活力与凋亡情况。

结果

成功构建出10例患者来源的胃癌类器官与肿瘤相关成纤维细胞，胃癌类器官表现出与原发肿瘤一致的形态学特征并在CK7、CEA与Ki-67的表达上具有相似性。肿瘤相关成纤维细胞呈现出典型的梭状及CD326

、CD45

、CD31

、α-SMA

、CD73

、CD90

、CD105

表型特征。与单独培养的类器官对照组比较，与肿瘤相关成纤维细胞共培养组的类器官形成数量更多、平均直径更长且总面积更大（

0.05）。在化疗药物5-氟尿嘧啶和奥沙利铂处理下，对照组的半数抑制浓度分别为10.66和3.26 μmol/L，而共培养组则分别为46.23和91.11 μmol/L；与类器官单独培养的对照组比较，共培养组中类器官的CellEvent™ Caspase 3/7 阳性凋亡细胞明显减少。

结论

与单独培养的类器官比较，肿瘤相关成纤维细胞共培养模型能更好地模拟体内肿瘤微环境的促瘤增殖与耐药效应。

Abstract

Objective‍　‍To construct an

in vitro

co-culture model of gastric cancer organoids and cancer-associated fibroblasts (CAFs)

and investigate the role of cancer-associated fibroblasts in the proliferation and chemotherapy resistance of gastric cancer organoi

ds. Methods‍　‍Tumor tissues from 12 gastric cancer patients undergoing surgical treatment in Department of General Surgery of Second Affiliated Hospital of Army Medical University from February 2023 to March 2024 were collected to construct gastric cancer organoids using 3D culture. HE staining was used to observe the morphology

and immunohistochemical assay was employed to determine the expression of cytokeratin CK7

carcinoembryonic antigen (CEA)

and proliferation marker Ki-67. After CAFs derived from the same patient were cultured

observed for their morphology under a light microscope

and detected for the phenotype by flow cytometry

the cells were co-cultured with gastric cancer organoids in a 1:1 ratio. Phase-contrast microscopy was applied to observe the growth of the organoids and analyze the number

average diameter

and total area. Then

organoids cultured alone served as the control group. After the control and co-culture groups were treated with chemotherapy drugs

5-fluorouracil and oxaliplatin

for 48 h

the viability and apoptosis of organoids were assessed with CellTiter-Glo

􀳏

3D assay and CellEvent™ Caspase 3/7 activity

respectively. Results‍　‍Gastric cancer organoids and CAFs were successfully established from 10 gastric cancer patient-derived samples. The gastric cancer organoids exhibited morphological characteristics consistent with the corresponding primary tumors

and showed positive expression of CK7

CEA

and Ki-67. CAFs displayed typical spindle-shaped morphology and exhibited the phenotypic markers CD326

CD45

CD31

α-SMA

CD73

CD90

and CD105

. Compared to the organoids cultured alone

the organoids co-cultured with CAFs showed more formation of organoids

in larger average diameter

and taking larger total area (

0.05). After the treatment of 5-fluorouracil and oxaliplatin

the half-maximal inhibitory concentration (IC

) was 10.66 and 3.26 μmol/L

respectively in t

he control group

while was 46.23 and 91.11 μmol/L in the co-culture group. Additionally

the number of CellEvent™ Caspase 3/7 positive apoptotic cells was significantly less in the co-culture group than the control group.

Conclusion

Compared with individually cultured gastric cancer organoids

the co-culture model of gastric cancer organoids and CAFs better simulates the pro-tumor proliferation and drug resistance effects of

in vivo

tumor microenvironment.

关键词

Keywords

references

SMYTH E C ， NILSSON M ， GRABSCH H I ， et al . Gastric cancer ［J］. Lancet ， 2020 ， 396 （ 10251 ）： 635 - 648 . DOI： 10.1016/S0140-6736（20）31288-5 http://dx.doi.org/10.1016/S0140-6736（20）31288-5 .

SUNG H ， FERLAY J ， SIEGEL R L ， et al . Global cancer statistics 2020： GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries ［J］. CA Cancer J Clin ， 2021 ， 71 （ 3 ）： 209 - 249 . DOI： 10.3322/caac.21660 http://dx.doi.org/10.3322/caac.21660 .

SHITARA K ， RHA S Y ， WYRWICZ L S ， et al . Neoadjuvant and adjuvant pembrolizumab plus chemotherapy in locally advanced gastric or gastro-oesophageal cancer （KEYNOTE-585）： an interim analysis of the multicentre， double-blind， randomised phase 3 study ［J］. Lancet Oncol ， 2024 ， 25 （ 2 ）： 212 - 224 . DOI： 10.1016/S1470-2045（23）00541-7 http://dx.doi.org/10.1016/S1470-2045（23）00541-7 .

LI G Z ， DOHERTY G M ， WANG J P . Surgical management of gastric cancer： a review ［J］. JAMA Surg ， 2022 ， 157 （ 5 ）： 446 - 454 . DOI： 10.1001/jamasurg.2022.0182 http://dx.doi.org/10.1001/jamasurg.2022.0182 .

ZHAI J ， SHEN J J ， XIE G P ， et al . Cancer-associated fibroblasts-derived IL-8 mediates resistance to cisplatin in human gastric cancer ［J］. Cancer Lett ， 2019 ， 454 ： 37 - 43 . DOI： 10.1016/j.canlet.2019.04.002 http://dx.doi.org/10.1016/j.canlet.2019.04.002 .

XU H X ， LYU X D ， YI M ， et al . Organoid technology and applications in cancer research ［J］. J Hematol Oncol ， 2018 ， 11 （ 1 ）： 116 . DOI： 10.1186/s13045-018-0662-9 http://dx.doi.org/10.1186/s13045-018-0662-9 .

XU H X ， JIAO D C ， LIU A G ， et al . Tumor organoids： applications in cancer modeling and potentials in precision medicine ［J］. J Hematol Oncol ， 2022 ， 15 （ 1 ）： 58 . DOI： 10.1186/s13045-022-01278-4 http://dx.doi.org/10.1186/s13045-022-01278-4 .

YANG L ， YANG S ， LI X Y ， et al . Tumor organoids： from inception to future in cancer research ［J］. Cancer Lett ， 2019 ， 454 ： 120 - 133 . DOI： 10.1016/j.canlet.2019.04.005 http://dx.doi.org/10.1016/j.canlet.2019.04.005 .

YANG Y J ， KONG Y J ， CUI J L ， et al . Advances and applications of cancer organoids in drug screening and personalized medicine ［J］. Stem Cell Rev Rep ， 2024 ， 20 （ 5 ）： 1213 - 1226 . DOI： 10.1007/s12015-024-10714-6 http://dx.doi.org/10.1007/s12015-024-10714-6 .

ZHAO Y ， LI S R ， ZHU L F ， et al . Personalized drug screening using patient-derived organoid and its clinical relevance in gastric cancer ［J］. Cell Rep Med ， 2024 ， 5 （ 7 ）： 101627 . DOI： 10.1016/j.xcrm.2024.101627 http://dx.doi.org/10.1016/j.xcrm.2024.101627 .

VLACHOGIANNIS G ， HEDAYAT S ， VATSIOU A ， et al . Patient-derived organoids model treatment response of metastatic gastrointestinal cancers ［J］. Science ， 2018 ， 359 （ 6378 ）： 920 - 926 . DOI： 10.1126/science.aao2774 http://dx.doi.org/10.1126/science.aao2774 .

DE VISSER K E ， JOYCE J A . The evolving tumor microenvironment： from cancer initiation to metastatic outgrowth ［J］. Cancer Cell ， 2023 ， 41 （ 3 ）： 374 - 403 . DOI： 10.1016/j.ccell.2023.02.016 http://dx.doi.org/10.1016/j.ccell.2023.02.016 .

FIORI M E ， DI FRANCO S ， VILLANOVA L ， et al . Cancer-associated fibroblasts as abettors of tumor progression at the crossroads of EMT and therapy resistance ［J］. Mol Cancer ， 2019 ， 18 （ 1 ）： 70 . DOI： 10.1186/s12943-019-0994-2 http://dx.doi.org/10.1186/s12943-019-0994-2 .

JENKINS L ， JUNGWIRTH U ， AVGUSTINOVA A ， et al . Cancer-associated fibroblasts suppress CD8 + T-cell infiltration and confer resistance to immune-checkpoint blockade ［J］. Cancer Res ， 2022 ， 82 （ 16 ）： 2904 - 2917 . DOI： 10.1158/0008-5472.CAN-21-4141 http://dx.doi.org/10.1158/0008-5472.CAN-21-4141 .

SCHUTH S ， LE BLANC S ， KRIEGER T G ， et al . Patient-specific modeling of stroma-mediated chemoresistance of pancreatic cancer using a three-dimensional organoid-fibroblast co-culture system ［J］. J Exp Clin Cancer Res ， 2022 ， 41 （ 1 ）： 312 . DOI： 10.1186/s13046-022-02519-7 http://dx.doi.org/10.1186/s13046-022-02519-7 .

LIN J L ， LIN J X ， LIN G T ， et al . Global incidence and mortality trends of gastric cancer and predicted mortality of gastric cancer by 2035 ［J］. BMC Public Health ， 2024 ， 24 （ 1 ）： 1763 . DOI： 10.1186/s12889-024-19104-6 http://dx.doi.org/10.1186/s12889-024-19104-6 .

MORGAN E ， ARNOLD M ， CONSTANZA CAMARGO M ， et al . The current and future incidence and mortality of gastric cancer in 185 countries， 2020-2040： a population-based modelling study ［J］. EClinicalMedicine ， 2022 ， 47 ： 101404 . DOI： 10.1016/j.eclinm.2022.101404 http://dx.doi.org/10.1016/j.eclinm.2022.101404 .

HAN B F ， ZHENG R S ， ZENG H M ， et al . Cancer incidence and mortality in China， 2022 ［J］. J Natl Cancer Cent ， 2024 ， 4 （ 1 ）： 47 - 53 . DOI： 10.1016/j.jncc.2024.01.006 http://dx.doi.org/10.1016/j.jncc.2024.01.006 .

BEEHARRY M K ， ZHANG T Q ， LIU W T ， et al . Optimization of perioperative approaches for advanced and late stages of gastric cancer： clinical proposal based on literature evidence， personal experience， and ongoing trials and research ［J］. World J Surg Oncol ， 2020 ， 18 （ 1 ）： 51 . DOI： 10.1186/s12957-020-01819-6 http://dx.doi.org/10.1186/s12957-020-01819-6 .

LIN Z J ， LI G Q ， JIANG K ， et al . Cancer therapy resistance mediated by cancer-associated fibroblast-derived extracellular vesicles： biological mechanisms to clinical significance and implications ［J］. Mol Cancer ， 2024 ， 23 （ 1 ）： 191 . DOI： 10.1186/s12943-024-02106-8 http://dx.doi.org/10.1186/s12943-024-02106-8 .

HIDALGO M ， AMANT F ， BIANKIN A V ， et al . Patient-derived xenograft models： an emerging platform for translational cancer research ［J］. Cancer Discov ， 2014 ， 4 （ 9 ）： 998 - 1013 . DOI： 10.1158/2159-8290.CD-14-0001 http://dx.doi.org/10.1158/2159-8290.CD-14-0001 .

QU S Q ， XU R Y ， YI G Z ， et al . Patient-derived organoids in human cancer： a platform for fundamental research and precision medicine ［J］. Mol Biomed ， 2024 ， 5 （ 1 ）： 6 . DOI： 10.1186/s43556-023-00165-9 http://dx.doi.org/10.1186/s43556-023-00165-9 .

LIU J Y ， LI P F ， WANG L ， et al . Cancer-associated fibroblasts provide a stromal niche for liver cancer organoids that confers trophic effects and therapy resistance ［J］. Cell Mol Gastroenterol Hepatol ， 2021 ， 11 （ 2 ）： 407 - 431 . DOI： 10.1016/j.jcmgh.2020.09.003 http://dx.doi.org/10.1016/j.jcmgh.2020.09.003 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Diagnostic value of endoscopic LCPL sign for high-risk intestinal metaplasia in gastric mucosa

Effect of calumenin on metastasis and invasion of gastric cancer and prognosis of patients

Remodeling of enhancers in high-grade epithelial dysplasia of gastric mucosa and its effect on expression of proliferation-related gene CD24

Effective prevention, early diagnosis and precise treatment of gastric cancer

Related Author

ZHENG Xipeng

PAN Huilin

WU Linyu

HOU Yongying

LIU Qin

MA Qiang

QIN Xiuyuan

LIU Kaijun

Related Institution

First Clinical College of Chongqing Medical University

Department of Pathology， Army Medical Center of PLA/Daping Hospital of Third Military Medical University， Chongqing

Department of Gastroenterology， Chongqing Key Laboratory of Precise Prevention and Treatment of Digestive Malignancies

Department of Gastroenterology，Chongqing Key Laboratory of Precise Prevention and Treatment of Digestive Malignancies，Army Medical Center of PLA/Daping Hospital of Third Military Medical University

Department of General Surgery， First Affiliated Hospital， Army Medical University （Third Military Medical University）

⁰