Uzunhisarcıklı Ebru, Bozkurt Nuh Mehmet, Sağlam Ayşe
Faculty of Pharmacy, Pharmacology Department, Erciyes University, Kayseri, Turkey.
Pharmacology Department, Institute of Health Sciences, Erciyes University, Kayseri, Turkey.
Med Oncol. 2025 Jul 24;42(9):372. doi: 10.1007/s12032-025-02952-6.
Glioblastoma is the most common primary malignant brain tumor. It is aimed to elucidate the role of ROCK in the regulation of PI3K signaling inhibited by PTEN and to evaluate the effects of ROCK inhibitors Thiazovivin and GSK 429286 on glioblastoma as a new therapeutic strategy. U87 MG and L929 cell lines were used to research the cytotoxic effect of ROCK inhibitors Thiazovivin and GSK 429286. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and xCELLigence real-time cell analyzer were used to determine its effect on cell viability. To assess the impact of ROCK inhibitors on PTEN protein levels, PTEN expression in cell lysates was quantitatively determined in vitro using the PTEN ELISA Kit. Total protein concentration was measured using the BCA protein assay kit, p-Akt and PTEN protein level was determined by the western blot method. Showed that GSK 429286 and Thiazovivin significantly reduced U87 MG cell viability even at low concentrations. When IC values were examined, it was understood that THV (IC: 11.02 µM) showed a more potent cytotoxic effect at lower concentrations compared to GSK 429286 (IC: 89.58 µM). At 50 µM, Thiazovivin and GSK 429286 concentration caused a significant increase in PTEN activity compared to the control and Temozolomide groups, even at low concentrations. THV 50 μM significantly increased PTEN expression compared to control (p < 0.05). GSK 50 μM, THV 1 μM, and THV 10 μM groups showed significantly reduced p-Akt expression (p < 0.05). Considering the effects on healthy cells, Thiazovivin being more effective at low doses suggests it may be a more selective agent. Our findings indicate that ROCK inhibitors may have cytotoxic effects on glioblastoma cells by affecting cell death mechanisms via a PTEN-associated reduction in PI3K/AKT signaling.
胶质母细胞瘤是最常见的原发性恶性脑肿瘤。旨在阐明ROCK在PTEN抑制的PI3K信号调节中的作用，并评估ROCK抑制剂噻唑维因和GSK 429286作为一种新的治疗策略对胶质母细胞瘤的影响。使用U87 MG和L929细胞系研究ROCK抑制剂噻唑维因和GSK 429286的细胞毒性作用。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法和xCELLigence实时细胞分析仪测定其对细胞活力的影响。为评估ROCK抑制剂对PTEN蛋白水平的影响，使用PTEN ELISA试剂盒在体外定量测定细胞裂解物中的PTEN表达。使用BCA蛋白测定试剂盒测量总蛋白浓度，通过蛋白质印迹法测定p-Akt和PTEN蛋白水平。结果显示，即使在低浓度下，GSK 429286和噻唑维因也能显著降低U87 MG细胞活力。检查IC值时发现，与GSK 429286(IC: 89.58 μM)相比，THV(IC: 11.02 μM)在较低浓度下显示出更强的细胞毒性作用。在50 μM时，即使在低浓度下，噻唑维因和GSK 429286浓度与对照组和替莫唑胺组相比也导致PTEN活性显著增加。与对照组相比，THV 50 μM显著增加PTEN表达(p < 0.05)。GSK 50 μM、THV 1 μM和THV 10 μM组显示p-Akt表达显著降低(p < 0.05)。考虑到对健康细胞的影响，噻唑维因在低剂量时更有效表明它可能是一种更具选择性的药物。我们的研究结果表明，ROCK抑制剂可能通过PTEN相关的PI3K/AKT信号减少影响细胞死亡机制，从而对胶质母细胞瘤细胞产生细胞毒性作用。
