Niwa Hidetomo, Furukawa Ken-Ichi, Seya Kazuhiko, Hirota Kazuyoshi
Department of Anesthesiology, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562, Japan.
Department of Pharmacology, Hirosaki University Graduate School of Medicine, Hirosaki, Aomori 036-8562, Japan.
Oncol Lett. 2017 Oct;14(4):4911-4917. doi: 10.3892/ol.2017.6806. Epub 2017 Aug 24.
The present study investigated the effects of N-methyl-D-aspartate receptor (NMDAR) antagonist ketamine, on the growth of gliomas. To analyze the effects of ketamine treatment, rat C6 glioma cells arising from astrocytes, and RNB cells representing non-malignant astrocytes, were examined. In ketamine-treated C6 cells, the gene expression changes associated with cell proliferation following ketamine treatment were evaluated using a cDNA microarray. A cell proliferation assay was performed to analyze the dose-dependent proliferation of C6 glioma and RNB cells following culture (72 h) with ketamine treatment (0-100 µM). Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays were performed following cell incubation with/without ketamine, to confirm if the ketamine-induced cell death of C6 glioma and RNB cells were due to apoptosis. In addition, cell proliferation and TUNEL assays were performed following cell incubations with a selective NMDAR antagonist, D-2-amino-5-phosphonovaleric acid (D-AP5). Analysis of the cDNA microarray indicated that the growth of C6 glioma cells were suppressed by the effects of ketamine. Furthermore, results of the proliferation assay confirmed that ketamine treatment inhibited C6 cell proliferation, most notably at a dose of 30 µM (n=7, 66.4%; P<0.001). The TUNEL assay results revealed that ketamine induced an apoptotic effect on C6 glioma cells, with a significant effect on the rate of death observed at all tested concentrations (3, 10, 30 and 100 µM). Results of the aforementioned proliferation and TUNEL assay experiments were reproduced when ketamine was replaced with a selective NMDAR antagonist, D-AP5. However, the NMDARantagonist-induced effects were not observed in RNB cell cultures. Although it would be premature to apply the results from the present study to human cases, these results indicated that ketamine is an anesthetic candidate providing potential benefit for glioma resection.
本研究调查了N-甲基-D-天冬氨酸受体（NMDAR）拮抗剂氯胺酮对胶质瘤生长的影响。为分析氯胺酮治疗的效果，研究人员检测了源自星形胶质细胞的大鼠C6胶质瘤细胞以及代表非恶性星形胶质细胞的RNB细胞。在用氯胺酮处理的C6细胞中，使用cDNA微阵列评估氯胺酮处理后与细胞增殖相关的基因表达变化。进行细胞增殖试验，以分析氯胺酮处理（0-100μM）培养（72小时）后C6胶质瘤细胞和RNB细胞的剂量依赖性增殖。在用/不用氯胺酮孵育细胞后进行末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）试验，以确认氯胺酮诱导的C6胶质瘤细胞和RNB细胞死亡是否是由于凋亡所致。此外，在用选择性NMDAR拮抗剂D-2-氨基-5-磷酸戊酸（D-AP5）孵育细胞后进行细胞增殖和TUNEL试验。cDNA微阵列分析表明，氯胺酮的作用抑制了C6胶质瘤细胞的生长。此外，增殖试验结果证实，氯胺酮处理抑制了C6细胞增殖，最显著的是在30μM剂量时（n = 7，66.4%；P<0.001）。TUNEL试验结果显示，氯胺酮对C6胶质瘤细胞具有凋亡作用，在所有测试浓度（3、10、30和100μM）下观察到的死亡率均有显著影响。当用选择性NMDAR拮抗剂D-AP5替代氯胺酮时，上述增殖和TUNEL试验结果得以重现。然而，在RNB细胞培养物中未观察到NMDAR拮抗剂诱导的效应。尽管将本研究结果应用于人类病例还为时过早，但这些结果表明氯胺酮是一种可为胶质瘤切除术带来潜在益处的麻醉候选药物。
