Haas Bodo, Roth Inken, Säcker Luisa, Wos-Maganga Maria, Beltzig Lea, Kaina Bernd
Federal Institute for Drugs and Medical Devices (BfArM), Kurt-Georg-Kiesinger-Allee 3, 53175, Bonn, Germany.
Faculty of Applied Natural Sciences, TH Köln - University of Applied Sciences, Campus Platz 1, 51379, Leverkusen, Germany.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Mar 24. doi: 10.1007/s00210-025-03955-w.
Temozolomide (TMZ) concomitant with radiotherapy is the first-line treatment for glioblastoma. However, treatment resistance is frequently observed in patients. Cellular senescence (CSEN) induced by TMZ has been proposed to be one underlying mechanism resulting in resting cells, causing inflammation and possibly recurrences if senescent cells re-enter the cell cycle after treatment. Inhibition of the K channels human ether-à-go-go type 1 (Eag1) and human ether-à-go-go-related gene (hERG) has shown promising effects in several tumor types including glioblastoma through growth inhibition and induction of apoptosis. In the present study, we analyzed the impact of hERG/Eag1 inhibition on apoptosis and CSEN on its own and in combination with TMZ in a panel of human glioblastoma cell lines and primary glioblastoma cells. hERG/Eag1 protein expression was determined by Western blotting and immunocytochemistry. Cytotoxicity of astemizole and terfenadine alone or in combination with TMZ was assessed by MTT assays. Apoptotic yields were determined by Annexin V/propidium iodide staining, and CSEN was quantified by determining SA-β-galactosidase levels through flow cytometry. We observed a similar protein expression of hERG and Eag1 in all glioblastoma cell lines and primary glioblastoma cells. Astemizole and terfenadine were cytotoxic in glioblastoma cells at low micromolar concentrations (5-10 µM range) through induction of apoptosis. In combination with TMZ, both drugs synergistically sensitized glioblastoma cells to TMZ-induced apoptosis. Moreover, astemizole reduced significantly the TMZ-induced CSEN level, indicating its impact on CSEN induction. Here, we show for the first time that blocking hERG/Eag1 channels in glioblastoma cells can relief TMZ-induced CSEN and synergistically ameliorates cytotoxicity through the induction of apoptosis.
替莫唑胺（TMZ）联合放疗是胶质母细胞瘤的一线治疗方案。然而，患者中经常出现治疗耐药情况。有人提出，TMZ诱导的细胞衰老（CSEN）是导致细胞静止的一种潜在机制，若衰老细胞在治疗后重新进入细胞周期，会引发炎症并可能导致复发。抑制钾通道人类醚 - 去极化型1（Eag1）和人类醚 - 去极化相关基因（hERG）在包括胶质母细胞瘤在内的多种肿瘤类型中，通过抑制生长和诱导凋亡显示出有前景的效果。在本研究中，我们分析了hERG/Eag1抑制对凋亡和CSEN的影响，以及其单独作用和与TMZ联合作用对一组人胶质母细胞瘤细胞系和原发性胶质母细胞瘤细胞的影响。通过蛋白质免疫印迹法和免疫细胞化学法测定hERG/Eag1蛋白表达。通过MTT法评估阿司咪唑和特非那定单独或与TMZ联合使用的细胞毒性。通过膜联蛋白V/碘化丙啶染色测定凋亡率，通过流式细胞术测定SA-β-半乳糖苷酶水平来定量CSEN。我们观察到所有胶质母细胞瘤细胞系和原发性胶质母细胞瘤细胞中hERG和Eag1的蛋白表达相似。阿司咪唑和特非那定在低微摩尔浓度（5 - 10 μM范围）下通过诱导凋亡对胶质母细胞瘤细胞具有细胞毒性。与TMZ联合使用时，两种药物均能协同增强胶质母细胞瘤细胞对TMZ诱导凋亡的敏感性。此外，阿司咪唑显著降低了TMZ诱导的CSEN水平，表明其对CSEN诱导有影响。在此，我们首次表明，阻断胶质母细胞瘤细胞中的hERG/Eag1通道可以缓解TMZ诱导的CSEN，并通过诱导凋亡协同改善细胞毒性。
