常春藤皂苷通过活性氧-p38/JNK通路调控食管鳞癌细胞迁移和凋亡的研究  

α-Hederin regulates migration and apoptosis by modulating the reactive oxygen species-p38/JNK pathway in esophageal squamous cell carcinoma cells

作　　者：任静宇 范玲玲[1] 段冷昕[1] 王梅林 REN Jing-yu;FAN Ling-ling;DUAN Leng-xin;WANG Mei-lin(School of Basic Medicine Sciences,Henan University of Science and Technology,Luoyang 471023,Henan Province,China;Department of Clinical Laboratory Diagnosis,The First Affiliated Hospital and Clinical Medical College,Henan University of Science and Technology,Luoyang 471023,Henan Province,China)

机构地区：[1]河南科技大学基础医学院,河南洛阳471023 [2]河南科技大学第一附属医院检验科,河南洛阳471023

出　　处：《中国临床药理学杂志》2023年第8期1147-1151,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河南省科技攻关基金资助项目(222102310362);河南科技大学博士启动基金资助项目(13480026)。

摘　　要：目的探究常春藤皂苷(α-Hederin)对人食管鳞状细胞癌KYSE-30细胞的增殖、迁移及凋亡的影响及其潜在机制。方法将KYSE-30细胞分为4组:空白对照组(常规培养)和低、中、高剂量组(5、10、20μmol·L^(-1)常春藤皂苷)。细胞用常春藤皂苷处理24 h,用划痕愈合实验、Transwell实验检测细胞的迁移能力,用Annexin V-FITC/PI双染流式细胞术检测细胞凋亡情况,用蛋白质印迹法检测磷酸化C-Jun氨基末端激酶蛋白(p-JNK)、磷酸化细胞外调节蛋白激酶蛋白(p-ERK)、磷酸化p38蛋白(p-p38)、B细胞淋巴瘤2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、裂解型多聚ADP核糖聚合酶(Cleaved PARP)、裂解型半胱氨酸天冬氨酸蛋白水解酶3(Cleaved caspase 3)的表达水平,用流式细胞仪检测细胞内活性氧(ROS)水平。结果空白对照组和低、中、高剂量组细胞24 h增殖抑制率分别为(2.07±0.35)%、(12.73±3.52)%、(27.37±5.50)%和(51.02±6.35)%;细胞凋亡率分别为(5.17±2.05)%、(10.73±2.28)%、(20.80±3.29)%和(37.23±3.65)%;空白对照组和低、中剂量组24 h;细胞迁移率分别为(40.02±4.02)%、(27.67±3.06)%和(13.33±4.73)%;ROS百分比分别为(4.17±1.19)%、(18.01±2.03)%和(35.50±3.66)%,以上指标,各剂量组与空白对照组比较,差异均有统计学意义(均P<0.05)。蛋白质印迹结果显示,常春藤皂苷可下调Bcl-2表达水平,上调Cleaved caspase 3、Cleaved PARP、Bax、p-JNK和p-p38表达水平。结论常春藤皂苷能够抑制KYSE-30细胞的增殖、迁移,促进其凋亡,机制可能与上调ROS-p38/JNK MAPK信号通路有关。Objective To explore the effects and potential mechanism ofα-Hederin on the proliferation,migration and apoptosis of human esophageal squamous cell carcinoma(ESCC)KYSE-30 cells.Methods KYSE-30 cells was divided into four groups:Control group(conventional culture),experimental-L,-M and-H groups(5,10,20μmol·L^(-1)α-Hederin).After 24 hours of treatment,the migration ability of cells was detected by scratch healing assay and Transwell assay;the apoptosis was detected by Annexin V-FITC/PI double staining flow cytometry;the protein expression levels of phosphorylated c-Jun N-terminal kinase(p-JNK),phosphorylated extracellular regulated protein kinase(p-ERK),phosphorylated p38 MAPK(p-p38),B-cell lym ph oma-2(Bcl-2),Bcl-2 associated X protein(Bax),Cleaved poly ADP-ribose polymerase(Cleaved PARP),Cleaved cysteinyl aspartate specific proteinase 3(Cleaved caspase 3)were detected by Western blot,and the level of intracellular reactive oxygen species(ROS)was detected by flow cytometry.Results The cell 24 h proliferation inhibition rates of control group and experimental-L,-M and-H groups were(2.07±0.35)%,(12.73±3.52)%,(27.37±5.50)%and(51.02±6.35)%,respectively;the apoptosis rates were(5.17±2.05)%,(10.73±2.28)%,(20.80±3.29)%and(37.23±3.65)%,respectively;the 24-hour cell migration rates of the control group and the experimental-L,-M groups were(40.02±24.02)%,(27.67±3.06)%and(13.33±4.73)%,respectively;the percentage of ROS in the control group,experimental-L,-M groups were(4.17±1.19)%,(18.01±2.03)%and(35.50±3.66)%,respectively.The above indexes were statistically significant between the administration group and the control group(all P<0.05).The results of Western blot showed thatα-Hederin could down-regulate the expression level of Bcl-2 and up-regulate the expression levels of Cleaved caspase 3,Cleaved PARP,Bax,p-JNK and p-p38.Conclusionα-Hederin inhibits the proliferation and migration of KYSE-30 cells,promotes cell apoptosis,the mechanism may be related to the up-regulation of ROS-p38/JNK MAPK signaling pa

关 键 词：常春藤皂苷 食管鳞癌 活性氧 迁移 凋亡 

分 类 号：R28[医药卫生—中药学]