Lung cancer is the leading cause of cancer mortality worldwide, with a median survival rate at 5 years of less than 20%. While molecular mapping aids in selecting appropriate therapies, it cannot predict personalized treatment response and long-term efficacy. For addressing these challenges, there is a great need for functional tests. Within this context, we developed patient-derived spheroids (PDS) from tumor and adjacent normal tissue to biomimic the respective tissue for assessing the personalized drug treatment response in vitro. Surgically resected lung specimens were used to generate spheroids using a two-step culture procedure. Flow cytometry and immune staining enabled the characterization of different cell populations resulting from the lung samples. PDS phenotype, cell proliferation and apoptosis were evaluated. Differential gene expression between tumor and adjacent normal tissue was analyzed via RT-qPCR. PDS drug sensitivity was assessed using a cell metabolic assay in response to two chemotherapeutic drug combinations. Cellular and molecular analysis revealed the proportion of epithelial cells, fibroblasts, and immune cells in the patients’ tissue samples. Subsequently, PDS models from tumor and normal lung were successfully established using the expanded epithelial cells. As a proof of concept, an analysis of the drug treatment using PDS of lung adenoid cystic carcinoma exhibited a dose-dependent effect in response to cisplatin/etoposide and cisplatin/paclitaxel. Our spheroid model of both tumor and non-tumor lung cells holds great promise for enhancing the treatment efficacy in the cancer patients.
肺癌是全球癌症死亡的首要原因，其中位五年生存率低于20%。尽管分子图谱分析有助于选择合适的治疗方案，但无法预测个体化治疗反应及长期疗效。为应对这些挑战，功能性检测手段的需求日益迫切。在此背景下，我们利用肿瘤组织及癌旁正常组织开发了患者来源球体模型，通过体外生物模拟相应组织来评估个体化药物治疗反应。采用手术切除的肺组织标本，通过两步培养法成功构建球体模型。流式细胞术与免疫染色技术实现了对肺组织样本中不同细胞群的特征分析。研究评估了患者来源球体的表型特征、细胞增殖与凋亡情况，并通过实时定量聚合酶链反应分析了肿瘤与癌旁正常组织的差异基因表达。采用细胞代谢检测法评估了球体模型对两种化疗药物组合的敏感性。细胞与分子分析揭示了患者组织样本中上皮细胞、成纤维细胞及免疫细胞的比例构成。随后，利用扩增的上皮细胞成功建立了肿瘤与正常肺组织的球体模型。概念验证阶段，对肺腺样囊性癌患者来源球体的药物处理分析显示，其对顺铂/依托泊苷和顺铂/紫杉醇的联合用药呈现剂量依赖性效应。我们建立的肿瘤与非肿瘤肺细胞球体模型，在提升癌症患者治疗效果方面展现出巨大潜力。
An Optimized Method to Culture Human Primary Lung Tumor Cell Spheroids
肿瘤(癌症)患者之家