Weng Xiong, Zhu Shun-Qin, Cui Hong-Juan
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
College of Biotechnology, Southwest University, Chongqing 400715, China.
Zhongguo Zhong Yao Za Zhi. 2018 Feb;43(4):772-778. doi: 10.19540/j.cnki.cjcmm.20171121.002.
Glioblastoma is a common brain tumor and the overall survival rate of the patients is very low, so it is an effective way to develop the potential chemotherapy or adjuvant chemotherapy drugs in glioblastoma treatment. As a well-known antimalarial drug, artesunate(ARTs) has clear side effects, and recently it has been reported to have antitumor effects, but rarely reported in glioblastoma. Different concentrations of ARTs were used to treat the glioblastoma cells, and then the inhibitory effect of ARTs on glioblastoma proliferation was detected by MTT assay; Ki67 immunofluorescence assay was used to detect the proliferation of cells; Soft agar experiment was used to explain the clonal formation abilities ; Flow Cytometry was used to detect the cell cycle; and Western blot assay was used to determine the expression of key cell cycle protein. MTT assay results indicated that ARTs-treated glioblastoma cell A172, U251, U87 were significantly inhibited in a time-and-dose dependent manner as compared to the control group(DMSO treatment group). Soft agar experiment showed that ARTs could significantly reduce the clonal formation ability of glioblastoma. Furthermore, Flow cytometry analysis showed that ARTs could obviously increase the cell proportion in G₀/G₁ phase and reduce the cell proportion in S phase. Western blot results showed that the expressions of cell cycle-related proteins CDK2, CDK4, cyclin D1 and cyclin B1 were all obviously down-regulated. Above all, ARTs may inhibit the proliferation of glioblastoma cells by arresting cell cycle in G₀/G₁ phase through down-regulating the expression of CDK2, CDK4, cyclin D1, cyclin B1. These results may not only provide a novel method for rediscovering and reusing ARTs but also provide a new potential drug for treating glioblastoma.
胶质母细胞瘤是一种常见的脑肿瘤，患者的总体生存率很低，因此开发潜在的化疗或辅助化疗药物是胶质母细胞瘤治疗的有效途径。青蒿琥酯（ARTs）作为一种著名的抗疟药物，具有明显的副作用，最近有报道称其具有抗肿瘤作用，但在胶质母细胞瘤中的报道很少。使用不同浓度的ARTs处理胶质母细胞瘤细胞，然后通过MTT法检测ARTs对胶质母细胞瘤增殖的抑制作用；采用Ki67免疫荧光法检测细胞增殖；软琼脂实验用于解释克隆形成能力；流式细胞术用于检测细胞周期；蛋白质免疫印迹法用于测定关键细胞周期蛋白的表达。MTT法结果表明，与对照组（二甲基亚砜处理组）相比，ARTs处理的胶质母细胞瘤细胞A172、U251、U87的增殖受到显著抑制，呈时间和剂量依赖性。软琼脂实验表明，ARTs可显著降低胶质母细胞瘤的克隆形成能力。此外，流式细胞术分析表明，ARTs可明显增加G₀/G₁期细胞比例，降低S期细胞比例。蛋白质免疫印迹结果表明，细胞周期相关蛋白CDK2、CDK4、细胞周期蛋白D1和细胞周期蛋白B1的表达均明显下调。综上所述，ARTs可能通过下调CDK2、CDK4、细胞周期蛋白D1、细胞周期蛋白B1的表达，使细胞周期停滞在G₀/G₁期，从而抑制胶质母细胞瘤细胞的增殖。这些结果不仅可为重新发现和重新利用ARTs提供一种新方法，也可为治疗胶质母细胞瘤提供一种新的潜在药物。
