摘要
目的:探讨斑蝥酸钠(sodium cantharidate,SCA)对人舌鳞癌CAL27细胞的抑制作用及相关机制。方法:利用不同浓度SCA预处理CAL27细胞后,采用CCK-8法分析细胞活力,划痕实验和Transwell法检测细胞迁移和侵袭能力,流式细胞仪检测细胞凋亡率,Western印迹法检测SCA对CAL27细胞p53及其磷酸化位点Ser33、Ser37、Ser46蛋白、抑凋亡蛋白B淋巴细胞瘤2(BCL-2)、促凋亡蛋白BCL-2相关X蛋白(BAX)和活化半胱氨酸天冬氨酸蛋白酶3(cleaved caspase 3)蛋白表达的影响。采用Graphpad Prism 9.0软件包对数据进行统计学分析。结果:与空白对照组相比,斑蝥酸钠组CAL27细胞增殖、迁移和侵袭能力显著降低,细胞凋亡率显著升高(P<0.01),呈剂量依赖性。随着SCA浓度增加,p53及其磷酸化位点Ser33、37、46蛋白表达显著上调(P<0.05或P<0.01),BCL-2蛋白表达下调,BAX蛋白表达显著上调,BCL-2/BAX比值显著降低,cleaved caspase 3蛋白表达显著上调(P<0.05或P<0.01)。结论:SCA能抑制人舌鳞癌CAL27细胞增殖、迁移和侵袭,并可能通过调控p53蛋白的磷酸化修饰,经BCL-2/BAX-caspase-3信号通路,诱导细胞凋亡。
PURPOSE:To investigate the inhibitory effect of sodium cantharidate(SCA)on human tongue squamous cell carcinoma CAL27 cells and its mechanism.METHODS:CAL27 cells were pretreated with different concentrations of SCA.Cell viability was analyzed by CCK-8 method.The migration and invasion of CAL27 cells were measured by scratch test and Transwell chamber,and the apoptosis rate was measured by flow cytometry.p53 protein and its phosphorylation sites Ser33,Ser37,Ser46,expression of BCL-2,BAX,and cleaved caspase 3 in CAL27 cells were detected by Western blot.Statistical analysis was performed with Graphpad Prism 9.0 software package.RESULTS:Compared with the blank control group,the proliferation,migration and invasion of CAL27 cells in sodium cantharidate group were significantly decreased,and the apoptosis rate was significantly increased(P<0.01)in a dose-dependent manner.The expression of p53 protein and its phosphorylation sites Ser33,Ser37,Ser46 protein was significantly up-regulated(P<0.05 or P<0.01).The expression of BCL-2 protein was down-regulated and the expression of BAX protein was significantly up-regulated(P<0.05 or P<0.01).The ratio of BCL-2/BAX was significantly decreased and the expression of cleaved caspase 3 protein was significantly up-regulated(P<0.05 or P<0.01).CONCLUSIONS:SCA can inhibit the proliferation,migration and invasion of human tongue squamous cell carcinoma CAL27 cells.It also down-regulates the ratio of BCL-2/BAX and up-regulates the expression of cleaved caspase 3 protein by regulating the phosphorylation of p53 protein,which induces apoptosis.
作者
李欣然
陈霖
孟箭
LI Xin-ran;CHEN Lin;MENG Jian(Xuzhou Clinical College,Xuzhou Medical University,Xuzhou 221000;Department of Stomatology,Xuzhou Central Hospital,Xuzhou 221000,Jiangsu Province,China)
出处
《上海口腔医学》
CAS
2024年第3期229-234,共6页
Shanghai Journal of Stomatology
基金
国家口腔疾病临床医学研究中心开放课题(NCRCO-202101)
徐州市科技计划项目(KC21187)。
关键词
斑蝥酸钠
舌鳞癌
CAL
27细胞
凋亡
p53
磷酸化
Sodium cantharidate
Tongue squamous cell carcinoma
CAL27 cell
Apoptosis
p53
Phosphorylation
