Institute of Translational Medicine, Nanchang University, 1299 Xuefu Avenue, Nanchang, Jiangxi 330001, PR China.
Department of Thyroid Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, PR China.
Life Sci. 2021 Apr 1;270:119112. doi: 10.1016/j.lfs.2021.119112. Epub 2021 Jan 26.
Glioblastoma is one of the most invasive tumors of the central nervous system, and has a high degree of malignancy and poor prognosis. Harmine, an active ingredient extracted from perennial herbs, has been reported to have obvious antitumor effects on various tumors. However, the effects of harmine on glioblastoma growth remain unknown. We here explored the effects of harmine on glioblastoma and its underlying molecular mechanisms related to tumorigenesis.
CCK-8 and immunofluorescent assay were performed to measure anti-proliferative effect of harmine on U251-MG and U373-MG cells. Wound healing assay was performed to measure the effects of harmine on cell migration. qRT-PCR and western blot were performed to detect the protein/gene expression. BALB/c nude mice bearing U251-MG xenografts was used to measure the effects of harmine on the growth of glioblastoma in vivo.
Harmine treatment significantly suppressed the proliferation of U251-MG and U373-MG cells in a dose and time-dependent way. Mechanistically, harmine reduced the basal and EGF-enhanced the phosphorylation level of FAK and AKT. Moreover, harmine inhibited the cell viability of U251-MG and U373-MG cells by downregulating the phosphorylation of the FAK/AKT pathway. Besides, harmine significantly suppressed the migration of U251-MG cells by suppressing the expression of MMP2, MMP9 and VEGF. Subsequently, orthotopic xenograft models revealed that harmine treatment dramatically inhibited the growth of glioblastoma in vivo.
In conclusion, these results suggest that harmine suppresses the proliferation and migration of U251-MG and U373-MG cells by inhibiting the FAK/AKT signaling pathway. Our findings elucidate harmine could be a promising drug for glioblastoma therapy.
胶质母细胞瘤是中枢神经系统最具侵袭性的肿瘤之一，具有高度恶性和预后不良的特点。哈尔明，一种从多年生草本植物中提取的活性成分，已被报道对各种肿瘤具有明显的抗肿瘤作用。然而，哈尔明对胶质母细胞瘤生长的影响尚不清楚。我们在此探讨了哈尔明对胶质母细胞瘤的影响及其与肿瘤发生相关的潜在分子机制。
CCK-8 法和免疫荧光法检测哈尔明对 U251-MG 和 U373-MG 细胞的抗增殖作用。划痕愈合实验检测哈尔明对细胞迁移的影响。qRT-PCR 和 Western blot 检测蛋白/基因表达。BALB/c 裸鼠皮下接种 U251-MG 异种移植瘤，检测哈尔明对体内胶质母细胞瘤生长的影响。
哈尔明处理以剂量和时间依赖的方式显著抑制 U251-MG 和 U373-MG 细胞的增殖。在机制上，哈尔明降低了 FAK 和 AKT 的基础磷酸化水平和 EGF 增强的磷酸化水平。此外，哈尔明通过下调 FAK/AKT 通路的磷酸化来抑制 U251-MG 和 U373-MG 细胞的活力。此外，哈尔明通过抑制 MMP2、MMP9 和 VEGF 的表达，显著抑制 U251-MG 细胞的迁移。随后，原位异种移植模型显示，哈尔明处理显著抑制了体内胶质母细胞瘤的生长。
总之，这些结果表明，哈尔明通过抑制 FAK/AKT 信号通路抑制 U251-MG 和 U373-MG 细胞的增殖和迁移。我们的研究结果表明，哈尔明可能是治疗胶质母细胞瘤的一种有前途的药物。
