摘要
目的探讨芍药苷(Paeoniflorin)抑制子宫内膜癌进展的作用及机制。方法通过TargetNet和Swiss target prediction在线数据库预测芍药苷靶基因,利用UALCAN数据库分析芍药苷靶基因在子宫内膜癌中的表达,并利用TCGA数据库分析芍药苷靶基因在不同临床病理特征间的表达情况。体外培养子宫内膜癌细胞株HEC108和Ishikawa,将子宫内膜癌细胞分成Control组/Vector组(细胞不处理),AURKA组(转染AURKA质粒),芍药苷组,芍药苷+AURKA组(芍药苷处理细胞并转染AURKA质粒)。生物信息学预测芍药苷的靶基因。采用不同浓度芍药苷作用细胞,CCK⁃8实验检测细胞活力,Transwell实验检测细胞迁移和侵袭能力,酶联免疫检测仪检测细胞乳酸脱氢酶(lactate dehydrogenase,LDH)活性,裸鼠移植瘤实验检测芍药苷体内抑制细胞增殖,免疫组化检测增殖细胞抗原(Ki⁃67),Western blot检测细胞中AURKA、迁移相关蛋白以及EMT相关蛋白表达。结果生物信息学显示AURKA是芍药苷的作用靶点,在子宫内膜癌组织中高表达且与临床不良表型有关(P<0.05)。体外实验验证结果显示,芍药苷可抑制HEC108和Ishikawa细胞增殖、迁移和侵袭能力,并增加LDH活性,抑制Ki⁃67表达(均P<0.05),抑制细胞中迁移相关蛋白(MMP2、MMP9)表达水平以及调控EMT相关蛋白(Vimentin、N⁃cadherin、Snail、E⁃cadherin)的表达水平(均P<0.05)。芍药苷作用裸鼠后可抑制移植瘤体积和重量增长(均P<0.05)。芍药苷呈浓度和时间依赖性抑制细胞中AURKA蛋白表达(均P<0.05)。AURKA高表达促进细胞增殖并降低LDH活性(均P<0.05)。芍药苷作用细胞后可抑制AURKA表达,降低细胞活性并提高LDH活性(均P<0.05)。结论芍药苷可在体外抑制子宫内膜癌细胞增殖、迁移和侵袭并在体内抑制裸鼠移植瘤生长,作用机制可能是芍药苷通过下调AURKA表达而抑制子宫内膜癌发展进程。
Objective To investigate the effect of Paeoniflorin in inhibiting the progression of endometrial cancer and its mechanism.Methods The target gene of Paeoniflorin was predicted by TargetNet and Swiss target prediction online databases,the expression of Paeoniflorin target gene in endometrial cancer was analyzed by UALCAN database,and the expression of Paeoniflorin target gene in patients with different clinicopathological characteristics was analyzed by TCGA database.The endometrial cancer cell lines HEC108 and Ishikawa were cultured in vitro,and were divided into the Control group/Vector group(no treatment),the AURKA group(transfected with AURKA plasmid),the Paeoniflorin group and the Paeoniflorin+AURKA group(treated with Paeoniflorin and transfected with AURKA plasmid).The target gene of Paeoniflorin was predicted by bioinformatics.The cells were treated with Paeoniflorin at different concentrations.The cells activity was detected by CCK⁃8 assay.The cells migration and invasion were detected by Transwell assay.The activity of lactate dehydrogenase(LDH)was detected by the enzyme⁃linked immunosorbent assay.The cells proliferation inhibited by Paeoniflorin in vivo was detected by the transplanted tumor assay in nude mice.The proliferating cell antigen(Ki⁃67)was detected by immunohisto⁃chemistry.The expression of AURKA,migration⁃related proteins and epithelial mesenchymal transition(EMT)⁃related proteins were detected by Western Blot.Results Bioinformatics showed that AURKA was the target of Paeoniflorin,which was highly expressed in endometrial cancer tissues and related to clinical adverse phenotypes(P<0.05).The vitro experiment results showed that Paeoniflorin could inhibit cells proliferation,migration and invasion of HEC108 and Ishikawa cells,increase LDH activity,inhibit Ki⁃67 expression(all P<0.05),the levels of migration⁃related proteins(MMP2,MMP9)in cells,and regulate the levels of EMT⁃related proteins(Vimentin,N⁃cadherin,Snail,E⁃cadherin)in cells(all P<0.05).Paeoniflorin could inhibit the growth of volume and the weight of transplanted tumors in nude mice(all P<0.05),as well as the expression of AURKA protein in concentration⁃and time⁃dependent manners(all P<0.05).The high expression of AURKA could promote cells proliferation and reduce LDH activity(all P<0.05).Conclusions Paeoniflorin inhibited the expression of Proliferation,migration and invasion of endometrial cancer cells in vitro and the growth of transplanted tumor in nude mice in vivo.The mechanism may be that Paeoniflorin inhibits the development of endometrial cancer by down⁃regulating AURKA expression.
作者
肖慧
覃鸿恩
姚姿羽
周辉
马荣河
XIAO Hui;QIN Hongen;YAO Ziyu;ZHOU Hui;MA Ronghe(Department of Pharmacy,the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture;Department of Pharmacy,Minda Hospital Affiliated to Hubei Minzu University,Enshi 445000,China)
出处
《中国癌症防治杂志》
CAS
2023年第6期623-630,共8页
CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
基金
湖北省中医药科研项目(ZY2019Z014)。
关键词
子宫内膜癌
芍药苷
增殖
迁移
侵袭
Endometrial cancer
Paeoniflorin
Proliferation
Migration
Invasion
