青蒿琥酯通过阻滞细胞周期抑制胶质母细胞瘤细胞增殖  

Artesunate inhibits proliferation of glioblastoma cells by arresting cell cycle

作　　者：翁熊 祝顺琴[1,2] 崔红娟[2] WENG Xiong;ZHU Shun-qin;CUI Hong-juan(School of Life Sciences, Southwest University, Chongqing 400715, China;State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China;College of Biotechnology, Southwest University, Chongqing 400715, China)

机构地区：[1]西南大学生命科学学院,重庆400715 [2]西南大学家蚕基因组与生物学国家重点实验室,重庆400715 [3]西南大学生物技术学院,重庆400715

出　　处：《中国中药杂志》2018年第4期772-778,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(31501100);重庆市基础与前沿研究计划项目(cstc2015jcyjA10120);中央高校基本科研业务费(XDJK2015C129);西南大学博士基金项目(SWU115021);西南大学本科生科技创新基金项目(20163107004)

摘　　要：胶质母细胞瘤是一种常见的脑瘤，患者生存率极低，开发潜在的化疗或辅助化疗药物是一条有效的途径。青蒿琥酯（artesunate，ARTs）是一种常见的抗疟疾药物，其毒副作用清楚，有研究表明具有抗肿瘤作用，但在胶质母细胞瘤中少有报道。该研究通过对不同浓度的ARTs处理胶质母细胞瘤细胞，采用MTr法检测ARTs对胶质母细胞瘤增殖的抑制作用、Ki67免疫荧光法检测细胞的增殖水平、软琼脂克隆形成实验检测肿瘤细胞体外克隆形成能力、流式细胞术检测细胞周期、Westernblot法检测ARTs对周期蛋白表达的影响。MTT细胞增殖实验结果显示与对照组相比（DMSO处理组），ARTs对胶质瘤细胞增殖具有抑制作用，呈浓度和时间依赖性。免疫荧光结果表明ARTs使得Ki67的阳性细胞显著减少；软琼脂实验结果显示ARTs显著降低胶质母细胞瘤的体外克隆形成能力；流式细胞术结果证实ARTs能使胶质母细胞瘤细胞G0／G1显著性增加，s期细胞比率显著性减少；Westernblot研究发现周期相关蛋白CDK2，CDK4，cyclinD1，eyelinB1表达明显下调。由此推测ARTs可能通过下调CDK2，CDK4，cyclinD1，cyclinB1的表达引起细胞周期阻滞来抑制胶质瘤细胞增殖。该研究为ARlTs的“老药新用”提供了新的途径，为寻找潜在的胶质瘤药物提供了新的思路。Glioblastoma is a common brain tumor and the overall survival rate of the patients is very low, so it is an efteetive way to develop the potential chemotherapy or adjuvant chemotherapy drugs in glioblastoma treatment. As a well-known antimalarial drug, arte- sunate （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 glioblastema prolif- eration was detected by MTY assay; Ki67 immunotluorescenee assay was used to detect the proliferation of cells; Soft agar experiment was used to explain the clonal formation abilities in vitro ; 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 resuks 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 elonal formation ability of glioblastoma. Furthermore,Flow cytometry analysis showed that ARTs could obviously increase the cell proportion in Go/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 Go/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.

关 键 词：青蒿琥酯 胶质母细胞瘤 增殖 细胞周期 

分 类 号：R285[医药卫生—中药学]