秋水仙碱对胶质瘤细胞系化疗敏感性的影响  

Effect of colchicine on the sensitivity of glioma cells to temozolomide chemotherapy

作　　者：吴宇[1] 朱馨艺 江洪祥 陈谦学[1] WU Yu;ZHU Xin-yi;JIANG Hong-xiang;CHEN Qian-xue(Department of Neurosurgery,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院神经外科,武汉430060

出　　处：《中国临床神经外科杂志》2024年第2期97-104,共8页Chinese Journal of Clinical Neurosurgery

摘　　要：目的探讨秋水仙碱对胶质瘤细胞系化疗敏感性的影响及其机制。方法采用不同浓度的替莫唑胺(TMZ)处理人脑胶质瘤细胞系U87和A172细胞,构建TMZ耐药细胞系U87/TR和A172/TR,再使用秋水仙碱(10、30 ng/ml)干预1、2、3 d,采用细胞计数法、BrdU法和流式细胞术检测细胞活力、细胞增殖和细胞周期。生物信息学方法分析胶质瘤组织差异表达的微小RNA(miRNAs),并预测其靶基因;然后,采用qRT-PCR检测U-87/TR和A172/TR细胞中这些miRNAs的表达情况,调控miRNAs的表达以观察秋水仙碱效应的变化。结果秋水仙碱显著抑制U87/TR和A172/TR细胞的增殖活力(P<0.05),呈时间依赖性和剂量依赖性(P<0.05),而且显著阻滞U87/TR和A172/TR的有丝分裂,使细胞阻滞在G0/G1期。30ng/ml秋水仙碱显著增加TMZ对U87/TR和A172/TR细胞增殖的抑制效果(P<0.05),明显降低TMZ的IC50值(P<0.05)。生信分析显示,胶质瘤组织miR-330-3p、miR-491-5p、miR-6782-5p、miR-31-5p、miR-330-5p、miR-137和miR-433-3p呈差异表达,秋水仙碱明显上调U87/TR和A172/TR细胞miR-330-3p的表达(P<0.05)、明显抑制miR-330-3p靶基因ErbB的表达(P<0.05)。抑制miR-330-3p表达明显逆转秋水仙碱的生物学效应(P<0.05)。结论秋水仙碱可以抑制TMZ耐药的胶质瘤细胞的增殖,其机制可能涉及到miR-330-3p/ErbB信号通路的调节。Objective To investigate the effect of colchicine on the sensitivity of glioma cells to temozolomide(TMZ)chemotherapy and its mechanism.Methods Different concentrations of TMZ were used to treat human glioma cell lines U87 and A172,and to construct TMZ-resistant cell lines U87/TR and A172/TR.After colchicine(10,30 ng/ml)treatment for 1,2,and 3 days,cell viability,cell proliferation,and cell cycle were detected by cell counting,BrdU method,and flow cytometry,respectively.The differentially expressed microRNAs(miRNAs)in glioma tissues were analyzed by bioinformatics methods,and their target genes were predicted.Then,qRT-PCR was used to detect the expression of these miRNAs in U87/TR and A172/TR cells,and the expression of miRNAs was regulated to observe the changes of colchicine effects.Results Colchicine significantly inhibited the proliferation of U87/TR and A172/TR cells(P<0.05),with time-and dose-dependent manners(P<0.05).Colchicine significantly blocked mitosis of U87/TR and A172/TR cells,and induced cell cycle arrest in the G0/G1 Phase.Colchicine at the concentration of 30 ng/ml significantly increased the inhibitory effect of TMZ on proliferation of U87/TR and A172/TR cells(P<0.05),and significantly reduced the IC50 value of TMZ(P<0.05).Bioinformatics analysis showed that miR-330-3p,miR-491-5p,miR-6782-5p,miR-31-5p,miR-330-5p,miR-137,and miR-433-3p were differentially expressed in glioma tissues.Colchicine significantly upregulated the expression of miR-330-3p in U87/TR and A172/TR cells(P<0.05),and significantly inhibited the expression of miR-330-3p target gene ErbB(P<0.05).Inhibition of miR-330-3p significantly reversed the biological effects of colchicine(P<0.05).Conclusions Colchicine can inhibit the proliferation of TMZ-resistant glioma cells,and the mechanism may be involved in the regulation of miR-330-3p/ErbB signaling pathway.

关 键 词：胶质瘤 U87细胞 A172细胞 替莫唑胺 秋水仙碱 化疗敏感性 

分 类 号：R739.41[医药卫生—肿瘤]