Ozpak Lütfiye, Göker Bağca Bakiye, Biray Avci Çiğir, Gündüz Cumhur, Durmaz Burak
Department of Medical Biology, Faculty of Medicine, Sütçü İMam University, Kahramanmaraş, Turkey.
Department of Medical Biology, Faculty of Medicine, Aydın Adnan Menderes University, Aydın, Turkey.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Apr 11. doi: 10.1007/s00210-025-04135-6.
This study aimed to assess the molecular effects of the miR- 155 - 5p in modulating Temozolomide chemosensitivity, which was examined utilizing in silico and in vitro methods on brain cancer stem cells (BCSCs) that exhibit high differentiation, apoptosis avoidance, and a remarkable capacity for self-renewal as a mechanism of chemotherapeutic resistance. The hypothesis of the study is how miR- 155 - 5p modulates temozolomide chemosensitivity in brain cancer stem cells (BCSCs) and their differentiated counterparts (BSCs) and whether it influences the molecular mechanisms underlying tumour recurrence and treatment failure. By silencing miR- 155 - 5p using anti-miR oligonucleotides, the knockdown efficiency of miR- 155 - 5p was confirmed via quantitative qRT-PCR. The increased chemosensitivity effect of miR- 155 - 5p against temozolomide in BCSCs and BCSs was measured by cytotoxicity assay. Subsequently, all studied groups were evaluated by performing cell viability, cell cycle, and apoptosis tests. miRNA-mRNA-pathway interactions were analyzed using different bioinformatics tools and databases. Cell viability was significantly reduced by 42.9% in the combined treatment (anti-miR- 155 - 5p + temozolomide) group compared to the untreated group. Temozolomide treatment in combination with anti-miR- 155 - 5p showed a significant decrease in cell viability compared to temozolomide alone. In contrast, late apoptosis increased 2.10-fold (p < 0.0001), indicating a synergistic effect in promoting late apoptosis. Combining temozolomide and anti-miR- 155 - 5p inhibits DNA synthesis by causing G2 phase arrest. mRNAs targeted by miR- 155 - 5p and significantly down-regulated in glioblastoma included GABRA1, GABRB2, SCN1A, GRIN2A, and SGIP1. By survival analysis, low expression of SCN1A was associated with poor prognosis (p < 0.05; HR = 0.7), highlighting its potential prognostic role. The combination of temozolomide treatment with suppression of miR- 155 - 5p may provide a more effective and side-effect minimized brain cancer treatment strategy by reducing resistance.
本研究旨在评估miR-155-5p在调节替莫唑胺化疗敏感性方面的分子效应，采用计算机模拟和体外方法，在具有高分化、避免凋亡以及显著自我更新能力（作为化疗耐药机制）的脑癌干细胞（BCSCs）上进行检测。本研究的假设是miR-155-5p如何调节脑癌干细胞（BCSCs）及其分化对应物（BSCs）对替莫唑胺的化疗敏感性，以及它是否影响肿瘤复发和治疗失败的分子机制。通过使用抗miR寡核苷酸沉默miR-155-5p，经定量qRT-PCR确认了miR-155-5p的敲低效率。通过细胞毒性试验测定了miR-155-5p对BCSCs和BCSs中替莫唑胺化疗敏感性增加的作用。随后，通过进行细胞活力、细胞周期和凋亡试验对所有研究组进行评估。使用不同生物信息学工具和数据库分析miRNA-mRNA-信号通路相互作用。与未治疗组相比，联合治疗（抗miR-155-5p + 替莫唑胺）组的细胞活力显著降低了42.9%。与单独使用替莫唑胺相比，替莫唑胺与抗miR-155-5p联合治疗显示细胞活力显著降低。相反，晚期凋亡增加了2.10倍（p < 0.0001），表明在促进晚期凋亡方面具有协同作用。替莫唑胺与抗miR-155-5p联合使用通过导致G2期阻滞抑制DNA合成。在胶质母细胞瘤中，miR-155-5p靶向且显著下调的mRNA包括GABRA1、GABRB2、SCN1A、GRIN2A和SGIP1。通过生存分析，SCN1A低表达与预后不良相关（p < 0.05；HR = 0.7），突出了其潜在预后作用。替莫唑胺治疗与抑制miR-155-5p相结合，可能通过降低耐药性提供一种更有效且副作用最小化的脑癌治疗策略。
