紫草素联合替莫唑胺抑制神经胶质瘤细胞的侵袭迁移  

Shikonin combined with temozolomide inhibited the invasion and migration of glioma cells

作　　者：李波[1] 刘炳辉 杨明[1] 洪伟力 应勇[1] 陈旦龙 李永宁[3] Li Bo;Liu Binghui;Yang Ming;Hong Weili;Ying Yong;Chen Danlong;Li Yongning(Department of Neurosurgery,Wenzhou Medical University,Taizhou,Zhejiang 318020,China;Department of Pathology,Huangyan Hospital,Wenzhou Medical University,Taizhou,Zhejiang 318020,China;Department of Emergency,The First Affiliated Hospital of Dalian Medical University,Dalian,Liaoning 116021,China)

机构地区：[1]温州医科大学黄岩医院神经外科,浙江台州318020 [2]温州医科大学黄岩医院病理科,浙江台州318020 [3]大连医科大学附属第一医院急诊科,辽宁大连116021

出　　处：《中国微侵袭神经外科杂志》2025年第2期104-108,共5页Chinese Journal of Minimally Invasive Neurosurgery

基　　金：浙江省基础公益研究计划项目(编号:LGF22H090027);台州市科技计划项目(编号:24ywa28)。

摘　　要：目的研究紫草素联合替莫唑胺(temozolomide,TMZ)对GL261胶质瘤细胞侵袭迁移的影响与机制。方法取对数生长期的GL261胶质瘤细胞,通过CCK-8法检测紫草素及TMZ对细胞存活率的影响,筛选出最佳药物浓度。实验分为:对照组、TMZ组、TMZ+紫草素组(T+S组)。通过流式细胞术检测细胞内活性氧;划痕实验检测细胞迁移率;Western blot检测基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)、丙酮酸激酶M2型(pyruvate kinase M2,PKM2)蛋白的表达情况。结果CCK-8法筛选出最佳药物浓度TMZ为400μmol/L和紫草素为2.5μmol/L。与对照组、TMZ组比较,T+S组活性氧明显升高(P<0.05)。T+S组作用24 h后,细胞呈死亡漂浮状态,划痕面积变大,细胞不再迁移,肿瘤细胞呈坏死凋亡状态。与对照组比较,T+S组MMP2、PKM2蛋白表达降低(P<0.05)。结论紫草素联合TMZ下调胶质瘤细胞MMP2、PKM2蛋白表达并抑制侵袭迁移,可能与其诱导活性氧水平上调有关。Objective To investigate the effects and mechanisms of shikonin combined with temozolomide(TMZ)on the invasion and migration of GL261 glioma cells.Methods GL261 glioma cells in the logarithmic growth phase were used.The effects of shikonin and TMZ on cell viability were detected using the CCK-8 assay to identify the optimal drug concentrations.The experiment was divided into control group,TMZ group,and TMZ+shikonin group(T+S group).Intracellular reactive oxygen species(ROS)were detected by flow cytometry.Cell mobility was detected by scratch assay.The expressions of matrix metalloproteinases(MMP2)and pyruvate kinase M2(PKM2)were detected by Western blotting.Results The optimal drug concentrations were determined to be 400μmol/L for TMZ and 2.5μmol/L for shikonin by the CCK-8 assay.Compared with the control and TMZ group,the T+S group showed a significant increase in ROS levels(P<0.05).After 24 hours of treatment with T+S,the cells showed a dead floating state,the scratch area increased,the cells no longer migrated,and the tumor cells showed a necrosis and apoptosis state.Compared with the control group,the expression levels of MMP2 and PKM2 proteins in the T+S group were significantly decreased(P<0.05).Conclusions Shikonin combined with TMZ downregulates the expression of MMP2 and PKM2 proteins in glioma cells and inhibits their invasion and migration,which may be related to the upregulation of ROS levels induced by the combination treatment.

关 键 词：神经胶质瘤 替莫唑胺 紫草素 活性氧 侵袭迁移 

分 类 号：R730.264[医药卫生—肿瘤]