Cytotoxic effects of Hypericum perforatum on Glioblastoma Cells by Inducing Oxidative Stress, Autophagy and Apoptosis.

Mesut Mete, Ulkun Unsal, Fatih Çöllü, Işil Aydemir, Erdoğan Kocamaz, Mustafa Barutçuoğlu, Beyhan Gürcü, Müge Karakayali, Ibrahim Tuğlu
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Abstract

Aim: St. John\'s Wort Oil (JWO) has a sedative property and it is used traditionally for the treatment of depression, neuralgia and excitability. JWO has been shown to have anticancer activity via apoptosis in glioblastoma cells. However, information on whether JWO is effective on the autophagy mechanism in glioblastoma is still not known. So, the current study was the first to search the autophagy mechanism T98 glioma cells.

Material and methods: Three groups were created with T98 human glioblastoma cells; Group 1: T98 glioma cells with no treated (Control group). Group 2: T98 glioma cells treated with 3 µl/ml JWO. Group 3: T98 glioma cells treated with 6 µl/ml JWO. The cell proliferation, oxidative stress, types of cell death were studied at IC50 dose of JWO.

Results: The proliferation of glioma cells was inhibited in 5.296 µl/ml dose. JWO induced apoptosis in T98 glioma cells in comparison with the control and there was statistically significant difference (p 0.001). Apoptosis was analyzed via TUNEL method and results were checked by flow cytometry. We also investigated the effects of JWO on autophagy in T98 glioma cells by immunostaining LC3-II and MDC fluorescent stainings. The differences between JWO treated and control group were notably significant (p 0.001). The immunofluorescence staining resultsof LC3-II was confirmed by Western blotting analysis.

Conclusion: JWO seems to be an effective treatment agent for glioblastoma. Not only does it induce apoptosis via oxidative stress but also affects the autophagy. The use of JWO in combination with other treatment options may increase the efficacy of treatment.

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贯叶连翘通过诱导氧化应激、自噬和凋亡对胶质母细胞瘤细胞的细胞毒作用。
目的:圣约翰草油(JWO)具有镇静作用,传统上用于治疗抑郁症、神经痛和兴奋性。JWO已被证明通过胶质母细胞瘤细胞的凋亡具有抗癌活性。然而,关于JWO是否对胶质母细胞瘤的自噬机制有效的信息尚不清楚。因此,本研究首次探索了胶质瘤细胞的自噬机制。材料与方法:用T98人胶质母细胞瘤细胞制备三组;第一组:未处理的胶质瘤细胞T98(对照组)。第二组:用3µl/ml JWO处理T98胶质瘤细胞。第三组:用6µl/ml JWO处理T98胶质瘤细胞。研究了IC50剂量下的细胞增殖、氧化应激和细胞死亡类型。结果:5.296µl/ml剂量对胶质瘤细胞增殖有抑制作用。与对照组相比,JWO诱导T98胶质瘤细胞凋亡,差异有统计学意义(p < 0.001)。TUNEL法分析细胞凋亡,流式细胞术检测细胞凋亡结果。我们还通过免疫染色LC3-II和MDC荧光染色研究了JWO对T98胶质瘤细胞自噬的影响。JWO组与对照组比较差异有统计学意义(p < 0.001)。Western blotting分析证实LC3-II免疫荧光染色结果。结论:JWO是治疗胶质母细胞瘤的有效药物。它不仅通过氧化应激诱导细胞凋亡,而且影响细胞自噬。JWO与其他治疗方案联合使用可提高治疗效果。
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