Advancements in 3-Dimensional (3D) culture models for studying disease have increased significantly over the last two decades, but fully understanding how these models represent in vivo still requires further investigation. The current study investigated differences in gene expression between a baseline sample and that maintained on a tissue-on-chip perfusion device for up to 96 h, with and without clinically-relevant doses of irradiation, to allow differentiation of model and treatment effects. Tumour tissue samples from 7 Head and Neck Squamous Cell Carcinomas (HNSCC) patients were sub-divided and either fixed immediately upon excision or maintained in a tissue-on-chip device for 48 and 96 h, with or without 2 Gray (Gy) or 10 Gy irradiation. Gene expression was measured using an nCounter®PanCancer Progression Panel. Differentially expressed genes between pre- and post-ex vivo culture, and control and irradiated samples were identified using nSolver software (version 4.0). The secretome from the tumour-on-chip was analysed for the presence of cytokines using a Proteome Profiler™ platform. Significant numbers of genes both increased (n= 6 and 64) and decreased (n= 18 and 58) in expression in the tissue maintained on-chip for 48 and 96 h, respectively, compared to fresh tissue; however, the irradiation schedule chosen did not induce significant changes in gene expression or cytokine secretion. Although HNSCC tissue maintained ex vivo shows a decrease in a large proportion of altered genes, 25% and 53% (48 and 96 h) do show increased expression, suggesting that the tissue remains functional. Irradiation of tumour tissue-on-chip needs to be conducted for longer time periods for specific gene changes to be observed, but we have shown, for the first time, the feasibility of using this perfusion platform for studying the genomic response of HNSCC tissue biopsies.
过去二十年间，用于疾病研究的三维（3D）培养模型技术取得显著进展，但这些模型如何准确反映体内真实情况仍需深入探究。本研究通过对比基线样本与在组织芯片灌注装置中培养长达96小时的样本（在有无临床相关剂量辐射条件下）的基因表达差异，以区分模型效应与处理效应。研究将7例头颈部鳞状细胞癌（HNSCC）患者的肿瘤组织样本进行分割处理：部分在切除后立即固定，其余置于组织芯片装置中培养48小时或96小时，并分别施加0、2戈瑞（Gy）或10 Gy的辐射干预。采用nCounter® PanCancer Progression Panel检测基因表达，使用nSolver软件（4.0版）鉴定离体培养前后样本及辐射处理组与对照组的差异表达基因。通过Proteome Profiler™平台分析芯片培养肿瘤组织的分泌蛋白质组中细胞因子表达情况。结果显示：与新鲜组织相比，芯片培养48小时与96小时的组织中分别出现显著上调基因（6个和64个）与下调基因（18个和58个）；但所选辐射方案未引起基因表达或细胞因子分泌的显著变化。尽管离体培养的HNSCC组织大部分差异基因表达下调，但仍有25%（48小时组）和53%（96小时组）的基因表达上调，表明组织保持生理功能。为观测特定基因变化，芯片肿瘤组织的辐射实验需延长处理时间。本研究首次证实了利用该灌注平台研究HNSCC组织活检样本基因组学反应的可行性。
肿瘤(癌症)患者之家