We aimed to identify the critical molecular pathways altered upon tumor stroma interactions in retinoblastoma (RB). In vitro 2 D cocultures of RB tumor cells (Weri-Rb-1 and NCC-RbC-51) with primary bone marrow stromal cells (BMSC) was established. Global gene expression patterns in coculture samples were assessed using Affymetrix Prime view human gene chip microarray and followed with bioinformatics analyses. Key upregulated genes from Weri-Rb-1 + BMSC and NCC-RbC-51 + BMSC coculture were validated using qRT-PCR to ascertain their role in RB progression. Whole genome microarray experiments identified significant (P ≤ 0.05, 1.1 log 2 FC) transcriptome level changes induced upon coculture of RB cells with BMSC. A total of 1155 genes were downregulated and 1083 upregulated in Weri-Rb-1 + BMSC coculture. Similarly, 1865 genes showed downregulation and 1644 genes were upregulation in NCC-RbC-51 + BMSC coculture. The upregulated genes were significantly associated with pathways of focal adhesion, PI3K-Akt signalling, ECM-receptor interaction, JAK-STAT, TGF-β signalling thus contributing to RB progression. Validation of key genes by qRT-PCR based revealed significant overexpression of IL8, IL6, MYC and SMAD3 in the case of Weri-Rb-1 + BMSC coculture and IL6 in the case of NCC-RbC-51 + BMSC coculture. The microarray expression study on in vitro RB coculture models revealed the pathways that could be involved in the progression of RB. The gene signature obtained when a growing tumor interacts with its microenvironment may provide new horizons for targeted therapy.

我们旨在鉴定在视网膜母细胞瘤（RB）中肿瘤基质相互作用后改变的关键分子途径。体外建立了RB肿瘤细胞（Weri-Rb-1和NCC-RbC-51）与原代骨髓基质细胞（BMSC）的二维共培养。使用Affymetrix Prime view人类基因芯片微阵列评估共培养样品中的全球基因表达模式，然后进行生物信息学分析。使用qRT-PCR验证了来自Weri-Rb-1 + BMSC和NCC-RbC-51 + BMSC共培养的关键上调基因，以确定它们在RB进程中的作用。全基因组微阵列实验确定了RB细胞与BMSC共培养后诱导的显着（P≤0.05，1.1 log 2 FC）转录组水平变化。在Weri-Rb-1 + BMSC共培养中，总共下调了1155个基因，上调了1083个基因。同样，在NCC-RbC-51 + BMSC共培养中，有1865个基因表达下调，有1644个基因表达上调。上调的基因与粘着，PI3K-Akt信号传导，ECM-受体相互作用，JAK-STAT，TGF-β信号传导通路显着相关，从而有助于RB的进展。通过基于qRT-PCR的关键基因的验证显示了蛋白的显着过表达在Weri-Rb-1 + BMSC共培养的情况下为IL8，IL6，MYC和SMAD3，在NCC-RbC-51 + BMSC共培养的情况下为IL6。体外RB共培养模型的微阵列表达研究揭示了可能与RB进程有关的途径。当生长中的肿瘤与其微环境相互作用时获得的基因标记可能为靶向治疗提供新的视野。