丹酚酸B对人鼻咽癌细胞株CNE-2增殖、凋亡的影响及机制研究  

Effects of salvianolic acid B on proliferation and apoptosis of human nasopharyngeal carcinoma cell line CNE-2 and its mechanism

作　　者：蒋学范[1] 张大勇[2] 胡未鸣[1] 浦立[1] 苏立众[1] JIANG Xuefan;ZHANG Dayong;HU Weiming;PU Li;SU Lizhong(Department of Otolaryngology,Center of Otolaryngology Head and Neck Sugery,Affiliated People's Hospital of Hangzhou Medical College,Zhejiang Provincial People's Hospital,Hangzhou,Zhejiang,310014,China;Department of Clinical Medicine,School of Medicine,Zhejiang University City College,Hangzhou,Zhejiang,310015,China)

机构地区：[1]浙江省人民医院,杭州医学院附属人民医院耳鼻咽喉科,耳鼻咽喉头颈外科中心,浙江杭州310014 [2]浙江大学城市学院医学院临床医学系,浙江杭州310015

出　　处：《中国耳鼻咽喉头颈外科》2021年第5期265-268,共4页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：浙江省中医药科学研究基金项目(2016ZA024);浙江省医药卫生研究基金项目(2016KYB024)。

摘　　要：目的研究丹酚酸B对人鼻咽癌CNE-2细胞增殖、凋亡的影响,并探讨其可能的分子机制。方法采用四甲基偶氮唑盐(MTT)法检测丹酚酸B对人鼻咽癌CNE-2细胞的半数抑制浓度(IC50);用IC50浓度的丹酚酸B处理人鼻咽癌CNE-2细胞后,采用MTT法检测各组细胞的增殖情况;在荧光显微镜下应用Hoechst33258荧光染色法观察细胞的凋亡情况;应用流式细胞仪分析细胞凋亡率情况;Western blot法检测细胞凋亡相关蛋白Bax、Bcl-2、Caspase-3、Caspase-9的表达水平。结果不同浓度的丹酚酸B对人鼻咽癌CNE-2细胞均有抑制作用,并呈浓度依赖性,与对照组相比差异具有统计学意义(P<0.05或P<0.01),IC50为(56.26±2.13)μg/ml,48 h最显著;Hoechst33258荧光染色发现典型的凋亡细胞核形态学变化;流式细胞仪检测显示实验组细胞的凋亡率高于对照组(P<0.01);与对照组相比,实验组Bax、Caspase-3、Caspase-9的蛋白表达显著增加,而Bcl-2的蛋白表达显著减少(P<0.01)。结论丹酚酸B能明显抑制人鼻咽癌CNE-2细胞增殖,促其凋亡,其机制可能与其阻滞细胞周期的转化并使凋亡相关蛋白表达的改变有关。OBJECTIVE To observe the effects and relative mechanisms of salvianolic acid B on the proliferation and apoptosis on human nasopharyngeal carcinoma CNE-2 cells.METHODS The median inhibitory concentration(IC50) of salvianolic acid B on human nasopharyngeal carcinoma CNE-2 cells was detected by Tetramethylazolium salt(MTT).Human nasopharyngeal carcinoma CNE-2 cells were treated with Salvianolic acid B at the concentration of IC50,and the proliferation of the cells in each group was detected by MTT.The apoptosis of the cells was observed by Hoechst33258 fluorescence staining under fluorescence microscope.The apoptosis rate was analyzed by flow cytometry(FCM).The expression levels of apoptosis related proteins Bax,Bcl-2,Caspase-3 and Caspase-9 were detected by Western blot.RESULTS Different concentrations of salvianolic acid B inhibited the human nasopharyngeal carcinoma CNE-2 cells,and in a concentration dependent manner,compared with the control group,the difference was statistically significant (P<0.05 or P<0.01),IC50 at (56.26±2.13)μg/ml,48 hours is the most significant.Typical morphological changes of apoptotic nuclei were found by Hoechst33258 fluorescence staining.Flow cytometry showed that the apoptosis rate of the experimental group was higher than that of the control group(P<0.01).Compared with the control group,the protein expression of Bax,Caspase-3 and Caspase-9 in the experimental group increased significantly,while the protein expression of Bcl-2 decreased significantly(P<0.01).CONCLUSION Salvianolic acid B can obviously inhibit the proliferation and induce the apoptosis of human nasopharyngeal carcinoma CNE-2 cells,and its mechanism may be related to the inhibition of cell cycle transformation and the change of expression of apoptosis related proteins.

关 键 词：鼻咽肿瘤 细胞增殖 细胞凋亡 丹酚酸B 

分 类 号：R285.5[医药卫生—中药学]