Minchenko Oleksandr H, Abramchuk Anastasiia I, Khikhlo Yevgen P, Sliusar Myroslava Y, Halkin Oleh V, Luzina Olha Y, Danilovsryi Serhiy V, Viletska Yuliia M, Minchenko Dmytro O
Department of Molecular Biology, Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv, Ukraine.
Department of Pediatrics, National Bohomolets Medical University, Kyiv, Ukraine.
Endocr Regul. 2025 Apr 21;59(1):48-56. doi: 10.2478/enr-2025-0007. Print 2025 Jan 1.
Endoplasmic reticulum (ER) stress and hypoxia are key factors for the effective growth of malignant tumors, including glioblastoma. The phosphoserine aminotransferase 1 (PSAT1) is an ER stress-responsive enzyme responsible for serine synthesis and necessary for tumor cell proliferation. The present study aims to investigate the regulation of the gene expression in U87MG glioblastoma cells and normal human astrocytes by ER stress and hypoxia depending on hydrocortisone, a native stress hormone used for co-treatment of glioblastoma and other malignant tumors. The U87MG glioblastoma cells and normal human astrocytes were used. Hypoxia was introduced by dimethyloxalylglycine. Tunicamycin was used for the induction of ER stress. Further, the cells were treated with hydrocortisone. RNA was extracted from cells after 4 h exposure to hydrocortisone, tunicamycin, and hypoxia. The expression level of the gene was studied by quantitative RT-PCR and normalized to ACTB mRNA. We found that treatment of normal human astrocytes with hydrocortisone resulted in a decreased expression of the gene, but its expression in glioblastoma cells was resistant to this hormone action. However, hypoxia did not significantly change the expression of the gene in normal astrocytes, but strongly modified the effect of hydrocortisone on this gene expression. At the same time, hypoxia increased the expression of the gene in glioblastoma cells independently of hydrocortisone. Tunicamycin decreased the expression of this gene in normal astrocytes, but increased it in glioblastoma cells. In addition, the impact of tunicamycin on gene expression was suppressed by hypoxia in both normal astrocytes and glioblastoma cells and by hydrocortisone only in normal astrocytes. At the same time, the combined effect of hypoxia and hydrocortisone greatly enhanced the expression of the gene in tunicamycin-treated normal astrocytes and especially glioblastoma cells. The results of this study showed that hydrocortisone differentially controls the regulation of gene expression by ER stress and hypoxia in normal astrocytes and glioblastoma cells and that the combined effect of hydrocortisone and hypoxia greatly enhanced gene expression in tunicamycin-treated cells.
内质网（ER）应激和缺氧是包括胶质母细胞瘤在内的恶性肿瘤有效生长的关键因素。磷酸丝氨酸转氨酶1（PSAT1）是一种内质网应激反应酶，负责丝氨酸合成，是肿瘤细胞增殖所必需的。本研究旨在探讨内质网应激和缺氧对U87MG胶质母细胞瘤细胞和正常人星形胶质细胞中该基因表达的调控作用，这取决于氢化可的松，一种用于胶质母细胞瘤和其他恶性肿瘤联合治疗的天然应激激素。使用了U87MG胶质母细胞瘤细胞和正常人星形胶质细胞。用二甲基草酰甘氨酸诱导缺氧。用衣霉素诱导内质网应激。此外，用氢化可的松处理细胞。在暴露于氢化可的松、衣霉素和缺氧4小时后从细胞中提取RNA。通过定量RT-PCR研究该基因的表达水平，并以ACTB mRNA进行标准化。我们发现，用氢化可的松处理正常人星形胶质细胞会导致该基因表达降低，但该基因在胶质母细胞瘤细胞中的表达对这种激素作用具有抗性。然而，缺氧并没有显著改变正常人星形胶质细胞中该基因的表达，但强烈改变了氢化可的松对该基因表达的影响。同时，缺氧在不依赖氢化可的松的情况下增加了胶质母细胞瘤细胞中该基因的表达。衣霉素降低了正常人星形胶质细胞中该基因的表达，但增加了胶质母细胞瘤细胞中该基因的表达。此外，衣霉素对该基因表达的影响在正常星形胶质细胞和胶质母细胞瘤细胞中均被缺氧抑制，而仅在正常星形胶质细胞中被氢化可的松抑制。同时，缺氧和氢化可的松的联合作用极大地增强了衣霉素处理的正常星形胶质细胞尤其是胶质母细胞瘤细胞中该基因的表达。本研究结果表明，氢化可的松在正常星形胶质细胞和胶质母细胞瘤细胞中通过内质网应激和缺氧对该基因表达的调控具有差异控制作用，并且氢化可的松和缺氧的联合作用极大地增强了衣霉素处理细胞中该基因的表达。
