Objective

Gliomas are the most prevalent type of malignant primary brain tumors. Despite the availability of several treatment modalities, these tumors have poor prognostic features. Aberrant Hedgehog (Hh) signaling has been found to be implicated in the development of numerous malignancies including gliomas. Naringenin appears to have anti-proliferative and anti-cancer properties. However, there is no report describing its effects via the Hh signaling pathway on the C6 glioblastoma cell line. The current study was set to examine the anti-cancer effects of naringenin on C6 cells in order to determine the effect of this compound on the Hh signaling pathway.

Methods

The anti-proliferative and apoptotic effects of naringenin against C6 and 3T3 fibroblast cells were measured by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and annexin-V/PI dual staining assay, respectively. The effect of naringenin on the migration of C6 cells was evaluated by the migration scratch assay. To assess the anti-cancer effect of naringenin on the Hh signaling pathway, the expression of Gli-1, Smo, and Sufu at protein levels in C6 cells was analyzed using western blotting.

Results

The obtained data indicated that naringenin exerted higher cytotoxicity against C6 cells (IC50 value of 114 ± 3.4 µg/mL) than normal 3T3 fibroblasts (IC50 value of 290 ± 7 µg/mL). Naringenin (114 µg/mL) also induced stronger apoptotic effects on C6 cells than 3T3 cells after 24 h of incubation. Furthermore, naringenin at a concentration of 114 µg/mL and a lower concentration of 60 µg/mL inhibited the migration of the C6 cell line. In addition, naringenin at a concentration of 114 µg/mL significantly decreased the expression of Gli-1 and Smo and elevated the expression of Sufu at the protein level in the C6 cell line.

Conclusion

These data represent that naringenin may have a potential effect on the management of the proliferation and metastasis of malignant gliomas by inhibiting the Hh signaling pathway.

中文翻译：

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柚皮素减弱 C6 胶质母细胞瘤细胞系的细胞活力和迁移：hedgehog 信号通路的可能作用

客观的

胶质瘤是最常见的恶性原发性脑肿瘤类型。尽管有几种治疗方式可供使用，但这些肿瘤的预后特征很差。已发现异常 Hedgehog (Hh) 信号传导与包括神经胶质瘤在内的许多恶性肿瘤的发展有关。柚皮素似乎具有抗增殖和抗癌特性。然而，没有报道描述其通过 Hh 信号通路对 C6 胶质母细胞瘤细胞系的影响。目前的研究旨在检查柚皮素对 C6 细胞的抗癌作用，以确定该化合物对 Hh 信号通路的影响。

方法

采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑 (MTT) 法和膜联蛋白-V/PI 双染色法测定柚皮素对 C6 和 3T3 成纤维细胞的抗增殖和凋亡作用分别测定。通过迁移划痕法评估柚皮素对C6细胞迁移的影响。为了评估柚皮素对 Hh 信号通路的抗癌作用，使用蛋白质印迹分析了 C6 细胞中 Gli-1、Smo 和 Sufu 在蛋白质水平上的表达。

结果

获得的数据表明，柚皮素对 C6 细胞的细胞毒性（IC50 值为 114 ± 3.4 µg/mL）比正常的 3T3 成纤维细胞（IC50 值为 290 ± 7 µg/mL）更高。与 3T3 细胞相比，Naringenin (114 µg/mL) 在孵育 24 小时后对 C6 细胞的凋亡作用也更强。此外，浓度为 114 µg/mL 和较低浓度的 60 µg/mL 的柚皮素抑制 C6 细胞系的迁移。此外，浓度为 114 µg/mL 的柚皮素显着降低了 C6 细胞系中 Gli-1 和 Smo 的表达，并提高了 Sufu 在蛋白质水平上的表达。

结论

这些数据表明，柚皮素可能通过抑制 Hh 信号通路对管理恶性胶质瘤的增殖和转移产生潜在影响。