五味子酯甲抑制头颈癌HN4细胞增殖和诱导凋亡的机制研究  

Schisantherin A inhibits cell proliferation and induces apoptosis of head and neck cancer HN4 cells

作　　者：曹祥燎 张路 钱峰[1,2] CAO Xiangliao;ZHANG Lu;QIAN Feng(Anhui Key Laboratory of Cancer Translational Medicine,Bengbu Medical College,Bengbu 233000,China;School of Pharmacy,Shanghai Jiaotong University)

机构地区：[1]蚌埠医学院癌症转化医学安徽省重点实验室,蚌埠233000 [2]上海交通大学药学院

出　　处：《山西医科大学学报》2021年第4期429-436,共8页Journal of Shanxi Medical University

基　　金：国家重点研发计划项目(2017YFC0908500);上海交通大学科技创新专项资金项目(19X160010005);国家自然科学基金项目(81973329,82073858,81773741)。

摘　　要：目的探讨五味子酯甲对人头颈癌HN4细胞增殖和凋亡的作用和分子机制。方法用不同浓度五味子酯甲(0,0.02,0.04,0.06,0.08,0.1μmol/L)处理人头颈癌HN4细胞7 d,检测五味子酯甲对细胞克隆性增殖的影响。采用CCK-8方法检测不同浓度五味子酯甲(0,0.05,0.1,0.5,1,2.5μmol/L)处理HN4细胞24 h的细胞存活率。用不同浓度的五味子酯甲(0,0.25,0.5,1μmol/L)处理HN4细胞24 h,采用PI染色的方法检测五味子酯甲对HN4细胞周期分布的影响,采用AnnexinⅤ-FITC/PI双染法检测五味子酯甲对HN4细胞凋亡的影响。用免疫印记法检测不同浓度五味子酯甲(0,0.25,0.5,1μmol/L)作用HN4细胞24 h后Cyclin B1、P21、DR5、DR4、cleaved Caspase-9、cleaved PARP蛋白相对表达水平。结果五味子酯甲剂量依赖性地减少HN4细胞克隆形成数目(P<0.05)。五味子酯甲剂量依赖性地降低HN4细胞存活率(P<0.05)。五味子酯甲浓度依赖性地诱导HN4细胞在S期和G2/M期阻滞和上调Cyclin B1和P21蛋白(P<0.05)。五味子酯甲浓度依赖性地诱导HN4细胞凋亡和上调DR5、DR4、cleaved Caspase-9、cleaved PARP蛋白(P<0.05)。结论五味子酯甲具有抑制细胞增殖、阻滞细胞周期和诱导细胞凋亡的作用,其机制可能与Cyclin B1、P21、DR5、DR4、cleaved Caspase-9、cleaved PARP蛋白的上调有关。Objective To investigate the effect of schisantherin A on cell proliferation and apoptosis of human head and neck cancer HN4 cells.Methods Head and neck cancer HN4 cells were incubated with different concentrations of schisantherin A at 0,0.02,0.04,0.06,0.08,0.1μmol/L for 7 d,and then the proliferation of HN4 cells was determined by colony formation assay.Cell survival rate was determined using cell counting kit-8 after treatment with 0,0.05,0.1,0.5,1,2.5μmol/L schisantherin A for 24 h.After treatment with 0,0.25,0.5 and 1μmol/L schisantherin A,PI staining assay was performed to detect the effect of schisantherin A on cell cycle,meanwhile,AnnexinⅤ-FITC/PI staining assay was used to evaluate the apoptosis of HN4 cells.Western blot assay was used to detect the protein expression levels of Cyclin B1,P21,DR5,DR4,cleaved Caspase-9,and cleaved PARP in HN4 cells after treatment with different concentrations of schisantherin A(0,0.25,0.5,1μmol/L)for 24 h.Results Schisantherin A dose-dependently inhibited the colony formation numbers of HN4 cells(P<0.05).Schisantherin A dose-dependently decreased the cell survival rate of HN4 cells(P<0.05).Schisantherin A induced the cell arrest at S phrase and G2/M phrase and up-regulated the expression of Cyclin B1 and P21 in a concentration-dependent manner(P<0.05).Schisantherin A induced HN4 cell apoptosis and up-regulated the expression levels of DR5,DR4,cleaved Caspase-9 and cleaved PARP proteins in a concentration-dependent manner(P<0.05).Conclusion Schisantherin A can inhibit the cell proliferation and induce the cell arrest and the cell apoptosis of HN4 cells by enhancing the expression of Cyclin B1,P21,DR5,DR4,cleaved Caspase-9 and cleaved PARP.

关 键 词：五味子酯甲 头颈癌 死亡受体途径 细胞凋亡 

分 类 号：R739.91[医药卫生—肿瘤]