Witusik-Perkowska Monika, Zakrzewska Magdalena, Jaskolski Dariusz J, Liberski Pawel P, Szemraj Janusz
Department of Medical Biochemistry, Medical University of Lodz, Lodz, Poland.
Department of Molecular Pathology and Neuropathology, Medical University of Lodz, Lodz, Poland.
Onco Targets Ther. 2019 May 20;12:3905-3918. doi: 10.2147/OTT.S190601. eCollection 2019.
The in vitro environment can influence not only the molecular background of glioblastoma drug-resistance and treatment efficiency, but also the mechanisms and pathways of cell death. Both crucial molecular pathways and the deregulation of miRNAs are thought to participate in tumor therapy-resistance. The aim of our study is to examine the potential influence of ex vivo conditions on the expression of miRNAs engaged in the machinery of tumor-drug resistance, since in vitro models are commonly used for testing new therapeutics. Glioblastoma-derived cells, cultured under three different sets of conditions, were used as experimental models in vitro. The expression of 84 miRNAs relevant to brain tumorigenesis was evaluated by multi-miRNA profiling for initial tumors and their corresponding cultures. Finally, the expression of selected miRNAs related to temozolomide-resistance (miR-125b, miR-130a, miR-21, miR-221, miR-222, miR-31, miR-149, miR-210, miR-181a) was assessed by real-time PCR for each tumor and neoplastic cells in cultures. Our results demonstrate significant discrepancies in the expression of several miRNAs between tumor cells in vivo and in vitro, with miR-130a, miR-221, miR-31, miR-21, miR-222, miR-210 being the most marked. Also differences were observed between particular models in vitro. The results of computational analysis revealed the interplay between examined miRNAs and their targets involved in processes of glioblastoma chemosensitivity, including the genes relevant to temozolomide response (, , , , ). The artificial environment may influence the selective proliferation of cell populations carrying specific patterns of miRNAs and/or the phenotype of neoplastic cells (eg differentiation) by the action of molecular events including miRNAs. These phenomena may influence the tumor-responsiveness to particular drugs, disturbing the evaluation of their efficacy in vitro, with unpredictable results caused by the interdependency of molecular pathways.
体外环境不仅会影响胶质母细胞瘤耐药性的分子背景和治疗效果，还会影响细胞死亡的机制和途径。关键分子途径和微小RNA（miRNA）的失调都被认为参与了肿瘤治疗耐药性。我们研究的目的是检测体外条件对参与肿瘤耐药机制的miRNA表达的潜在影响，因为体外模型常用于测试新疗法。在三种不同条件下培养的胶质母细胞瘤衍生细胞被用作体外实验模型。通过对原发性肿瘤及其相应培养物进行多miRNA分析，评估了84种与脑肿瘤发生相关的miRNA的表达。最后，通过实时聚合酶链反应（PCR）评估了每种肿瘤及其培养物中的肿瘤细胞中与替莫唑胺耐药相关的特定miRNA（miR-125b、miR-130a、miR-21、miR-221、miR-222、miR-31、miR-149、miR-210、miR-181a）的表达。我们的结果表明，体内和体外肿瘤细胞中几种miRNA的表达存在显著差异，其中miR-130a、miR-221、miR-31、miR-21、miR-222、miR-210最为明显。在体外的特定模型之间也观察到了差异。计算分析结果揭示了所检测的miRNA与其参与胶质母细胞瘤化学敏感性过程的靶标之间的相互作用，包括与替莫唑胺反应相关的基因（……）。人工环境可能通过包括miRNA在内的分子事件的作用，影响携带特定miRNA模式的细胞群体的选择性增殖和/或肿瘤细胞的表型（如分化）。这些现象可能会影响肿瘤对特定药物的反应性，干扰体外对其疗效的评估，分子途径的相互依赖性会导致不可预测的结果。
