Department of Neurological Surgery, The Second Affiliated Hospital of the Harbin Medical University, Harbin, 150001, China.
Department of Neurological Surgery, The Second Affiliated Hospital of the Harbin Medical University, Harbin, 150001, China.
Photodiagnosis Photodyn Ther. 2019 Jun;26:405-412. doi: 10.1016/j.pdpdt.2019.05.007. Epub 2019 May 11.
As a targeted therapeutic technique for glioma inhibition, photodynamic therapy (PDT) has gradually become a focus of basic research related to glioma treatment. The capacity of PDT to kill glioma cells involves varieties of pathways. In glioma cells, activated sodium-hydrogen exchanger isoform 1 (NHE1) can inhibit the cytotoxic effect of temozolomide (TMZ), promote cell migration and invasion, and inhibit cell apoptosis by changing the acid-base equilibrium. The purpose of our study was to explore if PDT combined with TMZ can effectively inhibit glioma cells by influencing NHE1 in vitro.
We analyzed the expression levels of proteins such as NHE1, ezrin, vimentin, Bcl-2, and Bax by Western blot analysis, we assessed the migration and invasion of rat C6 glioma cells by Transwell assay, and we evaluated C6 cell apoptosis in vitro by flow cytometry.
Western blot results indicated that NHE1, ezrin and vimentin were downregulated after cotreatment of C6 cells, and intracellular acidification was detected by a fluorometric intracellular pH assay. The migration and invasion capacities of C6 cells were significantly hindered after cotreatment, as shown by the Transwell assay. Experimental data also revealed a significant increase in cell apoptosis after cotreatment, as detected by flow cytometry; corresponding proapoptotic changes in Bcl-2, Bax and caspase-3 were also observed in vitro.
These results demonstrate that PDT combined with TMZ can inhibit C6 cell migration and invasion and promote mitochondrial-associated apoptosis by inhibiting NHE1. Therefore, this study provides supporting evidence for a potential method for the treatment of glioma.
光动力疗法（PDT）作为一种针对胶质瘤抑制的靶向治疗技术，已逐渐成为胶质瘤治疗相关基础研究的焦点。PDT 杀伤胶质瘤细胞的能力涉及多种途径。在胶质瘤细胞中，激活的钠离子-氢交换体亚型 1（NHE1）可通过改变酸碱平衡，抑制替莫唑胺（TMZ）的细胞毒性作用，促进细胞迁移和侵袭，并抑制细胞凋亡。本研究旨在探讨 PDT 联合 TMZ 是否能通过影响 NHE1 体外有效抑制胶质瘤细胞。
通过 Western blot 分析检测 NHE1、ezrin、vimentin、Bcl-2 和 Bax 等蛋白的表达水平，通过 Transwell 检测评估大鼠 C6 胶质瘤细胞的迁移和侵袭能力，通过流式细胞术评估 C6 细胞体外凋亡情况。
Western blot 结果表明，C6 细胞共处理后 NHE1、ezrin 和 vimentin 下调，通过荧光细胞内 pH 测定法检测到细胞内酸化。Transwell 检测显示 C6 细胞的迁移和侵袭能力明显受阻。流式细胞术检测还显示共处理后细胞凋亡明显增加；体外还观察到 Bcl-2、Bax 和 caspase-3 的促凋亡变化。
这些结果表明，PDT 联合 TMZ 通过抑制 NHE1 抑制 C6 细胞迁移和侵袭，并促进线粒体相关凋亡。因此，本研究为胶质瘤的治疗提供了一种潜在的方法。
