The impact of temozolomide and lonafarnib on the stemness marker expression of glioblastoma cells in multicellular spheroids

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 mu M lonafarnib (LNF), 100 mu M TMZ, or combination of 5 mu M LNF + 100 mu M 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.

Automated summary

This study investigates the impact of drug treatments on the expression of stemness markers in glioblastoma multiforme (GBM) cells using multicellular tumor spheroid (MCTS) models. The results show that drug treatment leads to a decrease in spheroid size and a significant increase in the expression of stemness markers. These findings highlight the importance of considering the effects of drug treatment on stemness marker expression when evaluating therapies.