摘要
目的:探讨黄芩素对人舌鳞状细胞癌(简称舌鳞癌)CAL27细胞增殖的影响,阐明其潜在的作用机制。方法:将对数生长期CAL27细胞分为对照组和不同浓度(12.5、25.0、50.0、100.0和200.0μmol·L^(-1))黄芩素组,采用结晶紫染色法观察各组细胞克隆形成情况,CCK-8法检测各组细胞增殖率,2’,7’-二氢二氯荧光素二乙酸酯(DCFH-DA)荧光探针检测各组细胞中活性氧(ROS)水平,罗丹明123(Rhodamine123)荧光探针检测各组细胞线粒体膜电位(MMP)水平。对数生长期CAL27细胞分为对照组和不同浓度(50、100和200μmol·L^(-1))黄芩素组,采用流式细胞术检测各组不同细胞周期细胞百分率和细胞凋亡率。对数生长期CAL27细胞分为对照组、不同浓度(50和100μmol·L^(-1))黄芩素组、N-乙酰半胱氨酸(NAC)组、50μmol·L^(-1)黄芩素+NAC组和100μmol·L^(-1)黄芩素+NAC组,采用DCFH-DA荧光探针和Rhodamine123荧光探针分别检测黄芩素与NAC联合作用后各组细胞中ROS和MMP水平。结果:结晶紫染色,与对照组比较,不同浓度黄芩素组细胞克隆形成数呈浓度依赖性减少,200μmol·L^(-1)黄芩素组细胞几乎无克隆形成。CCK-8法检测,与对照组比较,不同浓度黄芩素组细胞增殖率明显降低(P<0.05或P<0.01),且呈浓度依赖性。与对照组比较,不同浓度黄芩素组细胞中ROS水平明显升高(P<0.05),MMP水平明显降低(P<0.05)。与对照组比较,不同浓度黄芩素组细胞中S期细胞百分率明显升高(P<0.05),G0/G1期细胞百分率明显降低(P<0.05),细胞凋亡率明显升高(P<0.05)。黄芩素与NAC联合作用后,与对照组比较,50和100μmol·L^(-1)黄芩素组细胞中ROS水平明显升高(P<0.05),MMP水平明显降低(P<0.05);分别与50和100μmol·L^(-1)黄芩素组比较,50μmol·L^(-1)黄芩素+NAC组和100μmol·L^(-1)黄芩素+NAC组细胞中ROS水平明显降低(P<0.05),MMP水平明显升高(P<0.05)。结论:黄芩素可以通过激活线粒体氧化应激通路抑制CAL27细胞增殖。
Objective:To discuss the effect of baicalin on the proliferation of the human tongue squamous cell carcinoma(tongue squamous cell carcinoma)CAL27 cells,and to clarify its potential mechanism.Methods:The CAL27 cells at logarithmic growth phase were divided into control group and different concentrations(12.5,25.0,50.0,100.0,and 200.0μmol·L^(-1) )of baicalein groups,the clone formation of cells in various groups was observed by crystal violet staining;the proliferation rates of cells in various groups were detected by CCK-8 method;the levels of reactive oxygen species(ROS)in cells in vaious groups were detected by 2',7'-dichlorofluorescein diacetate(DCFH-DA)fluorescence probe;the mitochondrial membrane potential(MMP)of cells in various groups were detected by Rhodamine 123 fluorescence probe.The CAL27 cells at logarithmic growth phase were divided into control group and different concentrations(50,100,and 200μmol·L^(-1) )of baicalin groups.Flow cytometry was used to detect the percentages of the cells at different cell cycles and the apoptotic rates of cells in various groups.The CAL27 cells at logarithmic growth phase were divided into control group and different concentrations(50 and 100μmol·L^(-1) )of baicalein groups,N-neneneba acetylcysteine(NAC)group,50μmol·L^(-1) baicalin+NAC group,and 100μmol·L^(-1) baicalin+NAC group.The levels of ROS and MMP of cells in vraious groups were detected by DCFH-DA fluorescence probe and Rhodamine 123 fluorescence probe.Results:The crystal violet staining results showed that compared with control group,the numbers of clone formation of the cells in different concentrations of baicalin groups were decreased in a concentrationdependent manner,and there was almost no clone formation of the cells in 200μmol·L^(-1) baicalin group.The CCK-8 assay results showed that compared with control group,the proliferation rates of the cells in different concentrations of baicalin groups were significantly decreased in a concentration-dependent manner(P<0.05 or P<0.01).Compared with control group,the ROS levels of the cells in different concentrations of baicalin groups were significantly increased(P<0.05)and the MMP levels were significantly decreased(P<0.05).Compared with control group,the percentages of the cells at S phase in different concentrations of baicalin groups were significantly increased(P<0.05),the percentages of the cells at G0/G1 phase were significantly decreased(P<0.05),and the apoptotic rates were significantly increased(P<0.05).After the combination of baicalein and NAC,compared with control group,the ROS levels of the cells in 50 and 100μmol·L^(-1) baicalein groups were significantly increased(P<0.05)and the MMP levels were significantly decreased(P<0.05);compared with 50 and 100μmol·L^(-1) baicalein groups,the ROS levels of the cells in 50μmol·L^(-1) baicalein+NAC group and 100μmol·L^(-1) baicalein+NAC group were significantly decreased(P<0.05),and the MMP levels were significantly increased(P<0.05).Conclusion:Baicalein can inhibit the proliferation of the CAL27 cells by activating the mitochondrial oxidative stress pathway.
作者
史乃旭
郝苗
张天夫
赵柯林
黄子嫣
李春艳
王晓峰
SHI Naixu;HAO Miao;ZHANG Tianfu;ZHAO Kelin;HUANG Ziyan;LI Chunyan;WANG Xiaofeng(Department of Stomatology,China-Japan Union Hospital,Jilin University,Changchun 130033,China;Scientific Research Center,China-Japan Union Hospital,Jilin University,Changchun 130033,China;Department of Rheumatology and Immunology,China-Japan Union Hospital,Jilin University,Changchun 130033,China;Department of Oral and Maxillofacial Surgery,First Affiliated Hospital,Xinxiang Medical University,Xinxiang 453100,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2023年第4期985-993,共9页
Journal of Jilin University：Medicine Edition
基金
吉林省财政厅卫生科研人才专项项目(2021SCZ28)。
关键词
黄芩素
口腔鳞状细胞癌
细胞周期
活性氧
线粒体膜电位
Baicalien
Oral squamous cell carcinoma
Cell cycle
Reactive oxygen species
Mitochondrial membrane potential
