积雪草酸对前列腺癌细胞增殖及转化生长因子β1诱导的上皮间质转化的影响  

Effects of Asiatic Acid on Proliferation of Prostate Cancer Cells and Epithelial-mesenchymal Transition Induced by TGF-β1

作　　者：刘嘉 严宝飞 张景正 周兴宇 曾庆琪 LIU Jia;YAN Baofei;ZHANG Jingzheng;ZHOU Xingyu;ZENG Qingqi(Jiangsu Health Vocational College,Nanjing 211800 Jiangsu,China;The First Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Nanjing 210029 Jiangsu,China)

机构地区：[1]江苏卫生健康职业学院,江苏南京211800 [2]南京中医药大学第一附属医院,江苏南京210029

出　　处：《中药新药与临床药理》2022年第2期151-157,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：江苏省自然科学基金项目(BK20191498);国家中医药管理局名医验方评价与转化重点研究室开放课题(NZYJDMF-2020001);江苏省卫生健康委医学科研项目(重点项目,ZDB2020020);南京浦口区科技发展社会事业项目(S2020-14)。

摘　　要：目的观察积雪草酸对前列腺癌细胞DU145和PC-3增殖、迁移、侵袭的影响,并探讨其对上皮间质转化(EMT)的影响和潜在调控机制。方法体外培养DU145和PC-3细胞,以不同浓度积雪草酸干预后,采用MTT法检测其对细胞活力的影响;通过克隆形成实验检测其对细胞克隆形成能力的影响;通过细胞划痕实验、Transwell迁移及侵袭实验观察低浓度积雪草酸(10、20μmol·L^(-1))对转化生长因子β1(TGF-β1,5 ng·mL^(-1))诱导后DU145和PC-3细胞迁移、侵袭的影响;采用Western Blot法检测DU145细胞的血管内皮生长因子A(VEGFA)和EMT相关纤维连接蛋白(Fibronectin)、波形蛋白(Vimentin)、N-钙黏蛋白(N-Cadherin)、Snail及E-钙黏蛋白(E-Cadherin)的表达情况。结果积雪草酸可浓度及时间依赖性地抑制DU145和PC-3细胞增殖,干预24、48、72 h后的IC;值分别为29.10、17.52、11.53μmol·L^(-1)和27.91、21.43、14.82μmol·L^(-1)。积雪草酸(10、20μmol·L^(-1))能够明显抑制DU145及PC-3细胞克隆形成能力。与空白组比较,TGF-β1能够明显促进DU145和PC-3细胞的划痕愈合(P<0.01),促进DU145细胞的侵袭(P<0.05),上调DU145细胞内VEGFA蛋白和间质指标Fibronectin、Vimentin、N-Cadherin、Snail蛋白的表达(P<0.05,P<0.01),下调上皮指标E-Cadherin蛋白表达(P<0.05),进而诱导细胞EMT进程。与TGF-β1组比较,10、20μmol·L;浓度的积雪草酸能明显抑制TGF-β1诱导后的DU145、PC-3细胞划痕愈合及Transwell细胞迁移、侵袭(P<0.05,P<0.01),且呈现浓度依赖性;10μmol·L^(-1)浓度的积雪草酸能明显下调TGF-β1诱导的DU145细胞内的VEGFA、Fibronectin、Vimentin、N-Cadherin、Snail蛋白表达(P<0.05),上调E-Cadherin蛋白表达(P<0.05),进而逆转TGF-β1诱导的EMT进程。结论积雪草酸可抑制前列腺癌细胞DU145和PC-3增殖,并逆转TGF-β1诱导的前列腺癌细胞DU145的EMT进程,从而发挥其抗前列腺癌细胞侵袭、转移的作用。Objective To study the effects of asiatic acid(AA)on the proliferation,migration and invasion of human prostate cancer cells DU145 and PC-3,and to explore its effect on epithelial-mesenchymal transition(EMT)and the potential mechanism.Methods DU145 and PC-3 cells were cultured in vitro and treated with different concentrations of AA.The effects of AA on cell viability were detected by MTT assay.Colony forming assay was performed to assess the changes in colony forming compatibility of the cells.The effects of cell migration and invasion of AA(10μmol·L^(-1)and 20μmol·L^(-1))and TGF-β1(5 ng·mL^(-1))on DU145 and PC-3 cells were observed by cell scratching test,Transwell migration and invasion test.The protein expression of vascular endothelial growth factor A(VEGFA)and EMT related Fibronectin,Vimentin,N-cadherin,Snail and E-cadherin in DU145 cells were detected by Western Blot after intervened with AA and TGF-β1.Results AA significantly inhibited the proliferation of DU145 and PC-3 cells in a concentration-dependent and time-dependent manner,and the IC;values of AA were 29.10μmol·L^(-1)(24 h),17.52μmol·L^(-1)(48 h) and 11.53μmol·L^(-1)(72 h)for DU145 cells;27.91μmol·L^(-1)(24 h),21.43μmol·L^(-1)(48 h)and 14.82μmol·L^(-1)(72 h)for PC-3 cells.Compared with control group,TGF-β1 significantly promoted the scratched healing of DU145 and PC-3 cells(P<0.01),significantly promoted the expression of VEGFA and interstitial indexes such as Fibronectin,Vimentin,N-cadherin and Snail(P<0.05,P<0.01),significantly inhibited the expression of epithelial index such as E-cadherin in DU145cells(P<0.05),and then induced the process of EMT.Compared with TGF-β1 group,10μmol·L^(-1)and 20μmol·L^(-1)AA could significantly inhibit the scratched healing,migration and invasion of DU145 and PC-3 cells after TGF-β1 treatment(P<0.05,P<0.01)in a concentration-dependent manner.10μmol·L^(-1)AA significantly inhibited the expression of VEGFA,Fibronectin,Vimentin,N-cadherin and Snail(P<0.05),significantly promoted the expres

关 键 词：积雪草酸 前列腺癌 增殖 迁移 侵袭 上皮间质转化 转化生长因子Β1 

分 类 号：R285.5[医药卫生—中药学]