摘要
目的 探究冬凌草甲素对人软骨肉瘤细胞增殖、凋亡及p38 MAPK信号通路的调控作用.方法 体外培养人软骨肉瘤SW-1353 细胞,分为对照组(等量体积DMSO溶剂),5、10、15 μmol/L冬凌草甲素组(5、10、15 μmol/L冬凌草甲素)、10 μmol/L冬凌草甲素+抑制剂组(10 μmol/L冬凌草甲素+20 μmol/L p38 MAPK通路抑制剂SB203580)和阳性药物组(6 μmol/L阿霉素).用倒置显微镜、活细胞计数(CCK-8)、5-乙炔基-2′脱氧尿嘧啶核苷(EdU)、Hoechst 33258 染色、RT-qPCR及蛋白免疫印迹(WB)法对细胞形态、增殖活性、增殖率、凋亡情况、相关因子表达水平进行分析.结果 与对照组比较,实验组、10 μmol/L冬凌草甲素+抑制剂组和阳性药物组SW-1353 细胞增殖活性、增殖率、cyclin D1 及p-p38 MAPK表达水平显著降低,而细胞凋亡数量和caspase-3 表达水平增加,且浓度越高变化越显著(P<0.05),与 10 μmol/L冬凌草甲素组相比,抑制剂的加入使各指标变化更显著.结论 冬凌草甲素可通过抑制p38 MAPK通路抑制人软骨肉瘤SW-1353 细胞的增殖诱其凋亡.
Abstract
Objective To study effects of oridonin on proliferation,apoptosis and p38 MAPK signaling pathway of human chondrosarcoma cells.Methods In vitro experiments,human chondrosarcoma SW-1353 cells were divided into control group(equal volume of DMSO solven),5,10,15 μmol/L oridonin group(5,10,15 μmol/L oridonin),10 μmol/L oridonin+inhibitor group(10 μmol/L oridonin+20 μmol/L p38 MAPK signaling inhibitor SB203580)and positive drug group(6 μmol/L doxorubicin).The inverted microscope,CCK-8,5-acetylene-2′ deoxyuracil riboside(EdU)staining,Hoechst 33258 staining,RT-qPCR and western blot assays were used to detect cell morphology,viability,proliferation,apoptosis and related fator levels.Results Compared with control group,cell viability,proliferation rate and levels of cyclin D1 and p-p38 MAPK were significantly decreased in the 5,10,15 μmol/L oridonin group,10 μmol/L oridonin+inhibitor group and positive drug group,while cell apoptosis and caspase-3 level were significantly increased(P<0.05).Compared with the10 μmol/L oridonin group,the addition of inhibitors resulted in more significant changes in each index.Conclusion Oridonin could significantly inhibit the proliferation and promote apoptosis of SW-1353 cells by inhibiting of p38 MAPK signal transduction.
基金项目
秦皇岛市科学技术研究与发展计划项目(202301A266)
国家科技期刊平台
NSTL
万方数据