南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (2): 157-165.doi: 10.12122/j.issn.1673-4254.2023.02.01

• •    下一篇

体外重建宫颈癌的三维培养模型：基于人宫颈癌组织的去细胞支架

毛建英，杨文静，郭 和，董瑞丽，任丽芳，李树斌   

  1. 内蒙古自治区人民医院，内蒙古自治区 呼和浩特 010000
  • 出版日期:2023-02-20 发布日期:2023-03-16

A cervical cancer tissue-derived decellularized extracellular matrix scaffold for cervical cancer tissue reconstruction in vitro

MAO Jianying, YANG Wenjing, GUO He, DONG Ruili, REN Lifang, LI Shubin   

  1. Inner Mongolia Autonomous Region People's Hospital, Hohhot 010000, Inner Mongolia, China
  • Online:2023-02-20 Published:2023-03-16

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摘要： 目的 利用人宫颈癌组织制备去细胞化细胞外基质（ECM）支架组织，并评估该支架组织对3D培养模型中宫颈癌细胞生物学行为的影响。方法 采用十二烷基醚硫酸钠（SLES）溶液作为去细胞溶剂处理人宫颈癌组织，制备去细胞化ECM支架组织。通过病理学染色及生化含量分析等方法分析去细胞ECM支架组织的超微结构和关键成分胶原、糖胺聚糖、弹性蛋白及DNA含量，确定其最佳制备流程。将人宫颈癌SiHa细胞注射至去细胞化ECM支架组织中进行再细胞化，通过HE染色、免疫组织化学染色及分子生物学技术分析评估3D培养模型中及常规二维培养的SiHa细胞的迁移、增殖以及上皮间质转化（EMT），分析该支架组织中对宫颈癌细胞的生物学作用。用5-氟尿嘧啶（5-Fu）处理再细胞化3D培养模型后通过流式细胞术检测细胞凋亡，分析去细胞ECM支架组织对宫颈癌细胞抗药性的作用。结果 采用 SLES 溶液制备的宫颈癌组织去细胞ECM支架组织中，ECM的微环境结构及生物活性被有效地保留。在再细胞化的3D培养模型中，该ECM支架组织具有促进SiHa细胞增殖、迁移、EMT及抗药性的生物学作用。结论 本研究成功制备了基于宫颈癌组织的去细胞ECM支架组织，为构建宫颈癌体外3D研究模型奠定了基础。

关键词: 细胞外基质；去细胞支架；宫颈癌；肿瘤微环境

Abstract: Objective The prepare decellularized extracellular matrix (ECM) scaffold materials derived from human cervical carcinoma tissues for 3D culture of cervical carcinoma cells. Methods Fresh human cervical carcinoma tissues were treated with sodium lauryl ether sulfate (SLES) solution to prepare decellularized ECM scaffolds. The scaffolds were examined for ECM microstructure and residual contents of key ECM components (collagen, glycosaminoglycan, and elastin) and genetic materials by pathological staining and biochemical content analysis. In vitro 3D culture models were established by injecting cultured cervical cancer cells into the prepared ECM scaffolds. The cells in the recellularized scaffolds were compared with those in a conventional 2D culture system for cell behaviors including migration, proliferation and epithelial-mesenchymal transition (EMT) wsing HE staining, immunohistochemical staining and molecular biological technology analysis. Resistance to 5-fluorouracil (5-Fu) of the cells in the two culture systems was tested by analyzing the cell apoptosis rates via flow cytometry. Results SLES treatment effectively removed cells and genetic materials from human cervical carcinoma tissues but well preserved the microenvironment structure and biological activity of ECM. Compared with the 2D culture system, the 3D culture models significantly promoted proliferation, migration, EMT and 5-Fu resistance of human cervical cancer cells. Conclusion The decellularized ECM scaffolds prepared using human cervical carcinoma tissues provide the basis for construction of in vitro 3D culture models for human cervical cancer cells.

Key words: extracellular matrix; decellularized scaffolds; cervical carcinoma; tumor microenvironments