The impact of temozolomide and lonafarnib on the stemness marker expression of glioblastoma cells in multicellular spheroids
The impact of temozolomide and lonafarnib on the stemness marker expression of glioblastoma cells in multicellular spheroids
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DOI：
10.1002/btpr.3284
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发表时间：
2022-06
影响因子：
2.9
通讯作者：
Pinaki S. Nakod;Raghu Vamsi Kondapaneni;Brandon Edney;Yonghyun Kim;Shreyas S. Rao
中科院分区：
文献类型：
--
作者：
Pinaki S. Nakod;Raghu Vamsi Kondapaneni;Brandon Edney;Yonghyun Kim;Shreyas S. Rao
多形性胶质母细胞瘤（GBM）是一种高度恶性的脑肿瘤，预后不良。GBM微环境是高度异质性的，并且由许多细胞类型组成，包括星形胶质细胞和内皮细胞（EC）以及肿瘤细胞沿着，其负责对标准化疗药物如替莫唑胺（TMZ）的抗性增强。在这里，我们研究了药物治疗如何影响多细胞肿瘤球体（MCTS）模型中GBM细胞的干性标志物表达。将使用U87-MG GBM细胞、星形胶质细胞和/或EC构建的共培养和三培养MCTS培养7天。在第7天，加入5 μM洛那法尼（LNF）、100 μM TMZ或5 μM LNF + 100 μM TMZ的组合，并将MCTS再培养48小时。我们通过qRT-PCR和免疫染色评估了球体大小和干性标志物NESTIN、SOX 2、CD 133、NANOG和OCT 4的表达。在用LNF、TMZ或它们的组合（LNF + TMZ）处理48小时后，与未处理的对照相比，球状体尺寸减小。我们还观察到，与未处理的条件相比，在LNF + TMZ处理的条件下，大多数干性标志物的表达显著增加。这些结果表明，药物治疗可以影响MCTS模型中GBM细胞的干细胞标志物表达，在评估治疗时必须考虑这些方面。在未来，通过整合其他相关的细胞类型，我们可以进一步了解它们的串扰，最终导致新的治疗策略的开发。
Glioblastoma multiforme (GBM) is a highly malignant brain tumor with a poor prognosis. The GBM microenvironment is highly heterogeneous and is composed of many cell types including astrocytes and endothelial cells (ECs) along with tumor cells, which are responsible for heightened resistance to standard chemotherapeutic drugs such as Temozolomide (TMZ). Here, we investigated how drug treatments impact stemness marker expression of GBM cells in multicellular tumor spheroid (MCTS) models. Co‐ and tri‐culture MCTS constructed using U87‐MG GBM cells, astrocytes, and/or ECs were cultured for 7 days. At Day 7, 5 μM lonafarnib (LNF), 100 μM TMZ, or combination of 5 μM LNF + 100 μM TMZ was added and the MCTS were cultured for an additional 48 h. We assessed the spheroid sizes and expression of stemness markers‐ NESTIN, SOX2, CD133, NANOG, and OCT4‐ through qRT‐PCR and immunostaining. Following 48 h treatment with LNF, TMZ or their combination (LNF + TMZ), the spheroid sizes decreased compared to the untreated control. We also observed that the expression of most of the stemness markers significantly increased in the LNF + TMZ treated condition as compared to the untreated condition. These results indicate that drug treatment can influence the stemness marker expression of GBM cells in MCTS models and these aspects must be considered while evaluating therapies. In future, by incorporating other relevant cell types, we can further our understanding of their crosstalk, eventually leading to the development of new therapeutic strategies.