目的 分析确认小豆蔻明对人骨肉瘤细胞的抑制作用及相关机制。方法 分别用0、4、12和16 μmol·L-1小豆蔻明和体积分数0.08%的二甲基亚砜(DMSO)处理骨肉瘤细胞,结晶紫染色和四甲基偶氮唑蓝(MTT)实验检测细胞增殖;Hoechst染色和流式细胞术(flow cytometry,FCM)法检测细胞凋亡;划痕愈合和Transwell实验分别检测细胞迁移和侵袭;Western blot检测增殖、凋亡、迁移和侵袭相关蛋白以及Wnt/β-catenin信号通路相关蛋白的变化。结果 小豆蔻明抑制骨肉瘤细胞的增殖迁移和侵袭;抑制其增殖相关蛋白PCNA、迁移和侵袭相关蛋白基质金属蛋白酶-7(MMP-7)和波形蛋白、凋亡抑制蛋白Bcl-2的表达;促进其凋亡标志蛋白caspase 3和cleaved-caspase 3的表达;抑制Wnt/β-catenin信号关键分子β-catenin以及其下游靶分子cyclin D1和c-Myc的表达。结论 小豆蔻明可抑制骨肉瘤细胞的增殖、侵袭和迁移,促进其凋亡,其机制可能与干扰Wnt/β-catenin 信号通路的活化有关。
Abstract
OBJECTIVE To analyze and confirm the inhibitory effect of cardamonin(CAR) on human osteosarcoma(OS) cells and its related mechanism. METHODS Human osteosarcoma cells were treated with 0, 4, 12, 16 μmol·L-1 CAR and 0.08% DMSO, respectively. Cell proliferation was detected by crystal violet staining and MTT assay. Cell apoptosis was detected by Hoechst staining and flow cytometry(FCM). Cell migration ability was detected by scratch healing test. Cell invasion ability was detected by Transwell method. Western blot detects changes in cell proliferation, apoptosis, migration and invasion-associated proteins and Wnt/β-catenin signaling pathway-related proteins. RESULTS CAR inhibits proliferation, migration and invasion of osteosarcoma cells. CAR inhibits the expression of PCNA, MMP-7 and vimentin. CAR inhibits the expression of Bcl-2, promotes the expression of apoptotic markers caspase 3 and cleaved-caspase 3. CAR inhibits the expression of the key molecule β-catenin of Wnt/β-catenin signaling and its downstream target molecules cyclin D1 and c-Myc. CONCLUSION CAR can inhibit the proliferation, invasion and migration of osteosarcoma cells, but promote its apoptosis. Its molecular mechanism may be related to the interference of Wnt/beta-catenin signaling pathway activation.
关键词
小豆蔻明 /
骨肉瘤 /
抗肿瘤 /
Wnt/β-catenin 信号通路
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Key words
cardamonin /
osteosarcoma /
anti-cancer /
Wnt/beta-catenin signaling pathway
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中图分类号：
R965
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脚注
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基金
重庆市基础科学与前沿技术研究项目资助(cstc2017jcyjAX0039)
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