常春藤皂苷B对HGC-27胃癌细胞的抑制作用及基于生物信息学的机制探讨  

Inhibitory Effect of Hederasaponin B on Gastric Cancer HGC-27 Cells and Its Mechanism Based on Bioinformatics

作　　者：卫阳飞 王硕[1] 陈涛[1,2] 闫淑萍[1,2] 李佩佩 李洪梅 申诚 马玉梅[4] 宋志博 李艾静 李玉林 WEI Yangfei;WANG Shuo;CHEN Tao;YAN Shuping;LI Peipei;LI Hongmei;SHEN Cheng;MA Yumei;SONG Zhibo;LI Aijing;LI Yulin(Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining 810000,China;University of Chinese Academy of Sciences,Beijing 100000,China;Key Laboratory of Hexi Corridor Resources Utilization of Gansu,Hexi University,Zhangye 734000,China;Qinghai Institute of Health Sciences,Xining 810000,China)

机构地区：[1]中国科学院西北高原生物研究所,西宁810000 [2]中国科学院大学,北京100000 [3]河西学院甘肃省河西走廊特色资源利用重点实验室,甘肃张掖734000 [4]青海卫生职业技术学院,西宁810000

出　　处：《中国实验方剂学杂志》2022年第16期50-58,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82174052);甘肃省高等学校创新能力提升项目(2019A-102)。

摘　　要：目的:研究常春藤皂苷B对HGC-27胃癌细胞的抑制作用及机制。方法:通过噻唑蓝(MTT)比色法、苏木素-伊红(HE)染色、4',6-二脒基-2-苯基吲哚(DAPI)染色、平板克隆形成实验、划痕实验、流式细胞术测定细胞凋亡和细胞周期等,从不同角度研究了常春藤皂苷B对HGC-27胃癌细胞的抑制作用,进一步利用Pharm Mapper、UniProt、Swissdock、STRING和Metascape等在线平台,进行靶点筛选、基因注释、分子对接验证、蛋白质-蛋白质相互作用(PPI)网络构建、基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,探讨其作用机制。结果:与空白组比较,常春藤皂苷B(15、30、60、120μmol·L^(-1))可显著降低HGC-27胃癌细胞存活率(P<0.01),呈浓度和时间依赖性,常春藤皂苷B低于120μmol·L^(-1)时对正常细胞GES^(-1)无增殖抑制作用;与空白组比较,常春藤皂苷B(30、60、120μmol·L^(-1))可诱导HGC-27胃癌细胞胞质空泡的形成和细胞核变形固缩,抑制其迁移能力(P<0.01),诱导HGC-27胃癌细胞凋亡和细胞周期阻滞(P<0.05,P<0.01);当常春藤皂苷B 10、20、30μmol·L^(-1)时,对HGC-27胃癌细胞独立生存能力和增殖能力有抑制作用(P<0.01)。其作用靶点可能与驱动蛋白样蛋白KIF11、3',5'-环磷酸鸟苷特异性磷酸二酯酶、胱天蛋白酶-3、丝氨酸/苏氨酸蛋白激酶(Chk1)、原癌基因酪氨酸蛋白激酶、表皮细胞生长因子受体和丝裂原活化蛋白激酶(MAPK)8等相关,机制可能与癌症通路中的MAPK信号通路和幽门螺旋杆菌感染的上皮细胞信号通路中的黏附连接、局部连接、癌症中的蛋白聚糖通路等相关。结论:常春藤皂苷B对HGC-27胃癌细胞具有抑制作用,可能是多靶点多通路协同作用的结果。Objective:To investigate the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell and the mechanism.Method:Methyl thiazolyl tetrazolium(MTT)assay,hematoxylin-eosin(HE)staining,4',6-diamidino-2-phenylindote(DAPI)staining,colony formation assay,scratch assay,and flow cytometry were employed for the analysis of apoptosis and cell cycle.Thereby,the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell was investigated.Then the Pharm Mapper,UniProt,Swissdock,STRING,and Metascape were used for target screening,gene annotation,molecular docking,protein-protein interaction(PPI)network construction,Gene Ontology(GO)term and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis to explore the mechanism.Result:Hederasaponin B(15,30,60,120μmol·L^(-1))can significantly reduce the survival rate of HGC-27 cell(P<0.01)in a time-dependent and dosedependent manner compared with the blank group.It had no significant toxicity to normal GES^(-1) cell at concentration below 120μmol·L^(-1).Compared with the blank group,hederasaponin B(30,60,120μmol·L^(-1))induced cytoplasmic vacuolization,and nuclear deformation and karyopyknosis,inhibited the migration of HGC-27 cell(P<0.01),and brought about the apoptosis(P<0.05,P<0.01)and cell cycle arrest of HGC-27 cell(P<0.05,P<0.01).Hederasaponin B(10,20,30μmol·L^(-1))also suppressed the independent survival ability and proliferation ability of HGC-27 cell(P<0.01).The possible action targets were kinesin-like protein KIF11,cGMP-specific 3,5 cyclic phosphodiesterase,caspase-3,serine/threonine protein kinase Chk1,proto-oncogene tyrosine protein kinase,epidermal growth factor receptor,and mitogen-activated protein kinase(MAPK)8.The mechanism may be related to MAPK signaling pathway(pathways in cancer),adhesion connection,focal adhesion and proteoglycans in cancer(epithelial cell signaling pathways in Helicobacter pylori infection).Conclusion:Hederasaponin B exerts significant inhibitory effect on gastric cancer HGC-27 cell through multiple targets and multi

关 键 词：常春藤皂苷B HGC-27胃癌细胞 抑制作用 生物信息学 作用机制 

分 类 号：R22[医药卫生—中医基础理论] R2-031[医药卫生—中医学] R285.5R965