College of Life Science, Northeast Agricultural University, Harbin, China.
Key Laboratory of Neurosurgery, College of Heilongjiang Province, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
J Cell Biochem. 2019 Jan;120(1):622-633. doi: 10.1002/jcb.27420. Epub 2018 Sep 14.
Glioma, the most predominant primary malignant brain tumor, remains uncured due to the absence of effective treatments. Hence, it is imperative to develop successful therapeutic agents. This study aimed to explore the antitumor effects and mechanisms of ivermectin (IVM) in glioma cells in vitro and in vivo. The effects of IVM on cell viability, cell cycle arrest, apoptosis rate, and morphological characteristics were determined respectively by MTT assay/colony formation assay, flow cytometry, and transmission electron microscope. In addition, the expression levels of cycle-related and apoptosis-associated proteins were individually examined by Western blot analysis. Moreover, cell proliferation and apoptosis analyses were carried out by TUNEL, Ki-67, cleaved caspase-3, and cleaved caspase-9 immunostaining assay. Our results demonstrated that IVM has a potential dosage-dependent inhibition effect on the apoptosis rate of glioma cells. Meanwhile, the results also revealed that IVM induced apoptosis by increasing caspase-3 and caspase-9 activity, upregulating the expressions of p53 and Bax, downregulating Bcl-2, activating cleaved caspase-3 and cleaved caspase-9, and blocking cell cycle in G0/G1 phase by downregulating levels of CDK2, CDK4, CDK6, cyclin D1, and cyclin E. These findings suggest that IVM has an inhibition effect on the proliferation of glioma cells by triggering cell cycle arrest and inducing cell apoptosis in vitro and in vivo, and probably represents promising agent for treating glioma.
神经胶质瘤是最主要的原发性恶性脑肿瘤，由于缺乏有效的治疗方法，仍然无法治愈。因此，开发成功的治疗药物迫在眉睫。本研究旨在探讨伊维菌素（IVM）在体外和体内对神经胶质瘤细胞的抗肿瘤作用及其机制。通过 MTT 检测/集落形成实验、流式细胞术和透射电镜分别测定 IVM 对细胞活力、细胞周期阻滞、细胞凋亡率和形态特征的影响。此外，通过 Western blot 分析分别检测细胞周期相关和凋亡相关蛋白的表达水平。此外，通过 TUNEL、Ki-67、cleaved caspase-3 和 cleaved caspase-9 免疫染色检测进行细胞增殖和凋亡分析。我们的研究结果表明，IVM 对神经胶质瘤细胞的凋亡率具有潜在的剂量依赖性抑制作用。同时，结果还表明，IVM 通过增加 caspase-3 和 caspase-9 的活性、上调 p53 和 Bax 的表达、下调 Bcl-2、激活 cleaved caspase-3 和 cleaved caspase-9 以及通过下调 CDK2、CDK4、CDK6、cyclin D1 和 cyclin E 来阻断细胞周期在 G0/G1 期，诱导细胞凋亡。这些发现表明，IVM 通过触发细胞周期阻滞和诱导细胞凋亡，在体外和体内均对神经胶质瘤细胞的增殖具有抑制作用，可能是治疗神经胶质瘤的有前途的药物。
