Department of Histology and Embryology, Faculty of Medicine, Gaziantep Islam Science and Technology University, Türkiye;
Rom J Morphol Embryol. 2024 Jan-Mar;65(1):27-33. doi: 10.47162/RJME.65.1.04.
Cyclophosphamide (CP) is an alkylating chemotherapeutic agent commonly used in cancer treatments. In this study, we aimed to investigate the effects of 4-Hydroperoxy cyclophosphamide (4-HC), which is active form of CP, on glucose-regulated protein 78 (GRP78), activating transcription factor 6 (ATF6), phospho-protein kinase R (PKR)-like endoplasmic reticulum (ER) kinase (p-PERK), phospho-inositol-requiring enzyme 1 alpha (p-IRE1α), eukaryotic translation initiation factor 2 alpha (eIF2α), and caspase-3 messenger ribonucleic acids (mRNAs) and proteins that play roles in the ER stress pathway and apoptosis in U87 and T98 human glioblastoma cell lines. U87 and T98 human glioblastoma cell lines were divided into control and 4-HC-treated groups. Cell viability assay was used to detect the half maximal inhibitory concentration (IC50) for 24 hours of 4-HC. Immunocytochemistry and quantitative polymerase chain reaction (qPCR) methods were used to evaluate the levels of proteins and their mRNAs. The IC50 values of U87 and T98 cells were calculated as 15.67±0.58 μM and 19.92±1 μM, respectively. The levels of GRP78, ATF6, p-PERK, p-IRE1α, eIF2α, and caspase-3 protein expressions in the 4-HC-treated group compared to that in the control group. These increased protein expressions also were correlated with the mRNA levels. The ER stress signal pathway could be active in 4-HC-induced cell death. Further studies of ER-related stress mechanisms in anticancer treatment would be important for effective therapeutic strategies.
环磷酰胺（CP）是一种常用的癌症治疗烷化化疗药物。在这项研究中，我们旨在研究 4-羟过氧环磷酰胺（4-HC）对葡萄糖调节蛋白 78（GRP78）、激活转录因子 6（ATF6）、磷酸蛋白激酶 R（PKR）样内质网（ER）激酶（p-PERK）、磷酸肌醇需要酶 1α（p-IRE1α）、真核翻译起始因子 2α（eIF2α）和半胱天冬酶-3 信使 RNA（mRNA）和蛋白质的影响，这些蛋白质在 ER 应激途径和 U87 和 T98 人胶质母细胞瘤细胞系中的凋亡中发挥作用。将 U87 和 T98 人胶质母细胞瘤细胞系分为对照组和 4-HC 处理组。细胞活力测定法用于检测 4-HC 对 24 小时的半最大抑制浓度（IC50）。免疫细胞化学和定量聚合酶链反应（qPCR）方法用于评估蛋白质及其 mRNA 的水平。U87 和 T98 细胞的 IC50 值分别计算为 15.67±0.58 μM 和 19.92±1 μM。与对照组相比，4-HC 处理组的 GRP78、ATF6、p-PERK、p-IRE1α、eIF2α 和半胱天冬酶-3 蛋白表达水平升高。这些增加的蛋白表达也与 mRNA 水平相关。在 4-HC 诱导的细胞死亡中，内质网应激信号通路可能活跃。进一步研究 ER 相关应激机制在抗癌治疗中的作用对于制定有效的治疗策略非常重要。
