摘要
目的探讨骆驼蓬碱介导COX-2表达水平抑制骨肉瘤细胞增殖与凋亡的作用。方法体外培养人骨肉瘤细胞U2OS细胞系,随机分为对照组、研究组1、研究组2和研究组3,分别在0、5、10、20μmol/L浓度骆驼蓬碱条件下培养48 h。分别使用CCK-8法和流式细胞术检测细胞活力和细胞凋亡情况。并分别使用Western blot和实时荧光定量PCR(qRT-PCR)法检测COX-2、增殖和凋亡相关蛋白和mRNA的表达水平。结果在不同浓度骆驼蓬碱培养24、48 h后,三个研究组的细胞活力明显低于对照组(P <0. 05),而研究组2和研究组3的细胞活力均明显低于研究组1(P <0. 05)。在不同浓度骆驼蓬碱培养12、24、48 h后,三个研究组的细胞凋亡率明显高于对照组(P <0. 05),而研究组2和研究组3均明显高于研究组1(P <0. 05)。培养48 h后,研究组2的COX-2、Cyclin D1、PCNA、Bcl-2蛋白和mRNA的水平明显低于对照组,而cleaved caspase-3和Bax表达水平明显高于对照组(P <0. 05)。结论骆驼蓬碱具有抗骨肉瘤的作用,骆驼蓬碱可通过抑制COX-2的表达调节细胞周期相关蛋白和细胞凋亡相关蛋白的表达发挥抑制骨肉瘤细胞增殖和促进其凋亡的作用。
Objective To investigate the inhibitory effect of harmine on osteosarcoma cell proliferation and apoptosis by down regulating cyclooxygenase-2(COX-2)expression.Methods Human osteosarcoma cell line U20S was cultured in vitro and randomly divided into control group,study group I,study group 2 and study group 3.The cells were cultured in 0,5μmol/L,10 jxmol/L and 20μmol/L concentration of harmine for 48 hours.Cell counting kit-8(CCK-8)method and flow cytometry were used to detect cell viability and apoptosis.The expression level of COX-2,proliferation and apoptosis related proteins and mRNA were detected by Western blot and real-time quantitative polymerase chain reaction(RT-qPCR),respectively.Results After cultured with different concentrations of harmine for 12,24 hours and 48 hours,the cell viability of the three study groups were significantly lower than that of the control group(P<0.05),while that of the study groups 2 and 3 were significantly lower than that of the study group 1(P<0.05).The apoptosis rate of the three study groups were significantly higher than that of the control group(P<0.05),while that of the two groups were significantly higher than that of the study group 1(P<0.05).After 48 hours of culture,the levels of COX-2,cyclin Dl,proliferating cell nuclear antigen(PCNA),B-cell lymphoma-2(Bcl-2)protein and mRNA expression in study group 2 were significantly lower than those in control group,while the expression levels of cleaved caspase-3 and BCL2-Associated X(Bax)in study group 2 were significantly higher than those in control group(P<0.05).Conclusions Harmine can inhibit the proliferation and promote the apoptosis of osteosarcoma cells by inhibiting the expression of COX-2 and regulating the expression of cell cycle and apoptosis related protein.
作者
李钟陈
陈珀航
金建强
Li Zhongchen;Chen Cuhang;Jin Jianqiang(Department of Onhopaedics,Haining Hospital,Zhejiang People’s Hospital,Haining 314408,China;Department of Orthopaedics,Zhejiang People's Hospital,Hangzhou 3IOO14,China)
出处
《中国医师杂志》
CAS
2020年第2期207-210,215,共5页
Journal of Chinese Physician
基金
浙江省自然科学基金(LQ18H060004)。
