冬凌草甲素对结肠癌LOVO细胞增殖和EMT影响的实验研究  

The experimental study of oridonin on proliferation and EMT of colon cancer LOVO cells

作　　者：龙景培[1] 卜贺启 刘殿雷 Long Jingpei;Bu Heqi;Liu Dianlei(Department of General Surgery,the Women's Hospital,School of Medicine,Zhejiang University,Hangzhou 310006,China;Department of Coloproctological Surgery,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]浙江大学医学院附属妇产科医院外科,浙江杭州310006 [2]浙江省立同德医院肛肠科,浙江杭州310012

出　　处：《实用肿瘤杂志》2019年第4期309-315,共7页Journal of Practical Oncology

基　　金：浙江省医药卫生科技计划面上项目(2018KY325);浙江省自然科学基金项目(Y17H160187)

摘　　要：目的探讨冬凌草甲素(oridonin,ORI)对结肠癌细胞株LOVO细胞增殖和上皮间质转化(epithelialmesenchymal transition,EMT)的影响及可能的机制。方法不同药物浓度(0、2、4、8、12和16μg/m L)冬凌草甲素处理结肠癌LOVO细胞,不同时间点(24、48和72 h)采用CCK-8法观察细胞增殖情况,RT-PCR检测E-cadherin和Vimentin mRNA表达水平,Western blot检测磷酸化糖原合成酶激酶-3β(phospharylated glycogen synthase kinase-3β,p-GSK-3β)、β-catenin、E-cadherin和Vimentin蛋白表达水平。结果冬凌草甲素对LOVO细胞增殖抑制率呈时间和剂量依赖性(均P<0.05)。4、8和12μg/m L冬凌草甲素作用48 h,增加LOVO细胞E-cadherin基因表达量,减少Vimentin的基因表达量(均P<0.05),也增加LOVO细胞p-GSK-3β(Tyr216)蛋白表达量,减少p-GSK-3β(Ser9)蛋白表达量,同时下调β-catenin蛋白表达量(均P<0.05)。GSK-3β特定的抑制剂CHIR 2μmol/L作用1 h后,再加入12μg/m L冬凌草甲素共同作用24 h,能逆转冬凌草甲素对β-catenin、E-cadherin和Vimentin蛋白表达的影响(均P<0.05)。结论冬凌草甲素可能通过Wnt/β-catenin信号通路抑制结肠癌LOVO细胞的生长增殖和EMT过程。Objective To investigate the effect of oridonin on the proliferation and epithelial-mesenchymal transition(EMT)of colon cancer LOVO cell line and possible mechanisms.Methods CCK-8 method was used to observe the effect of(0,2,4,8,12,16μg/mL)oridonin on the proliferation of colon cancer LOVO cells at different time points(24,48 and 72 h);and the mRNA expressions of E-cadherin and Vimentin were detected by RT-PCR;and the protein expressions of phospharylated glycogen synthase kinase-3β(p-GSK-3β),β-catenin,E-cadherin and Vimentin were analyzed by Western blot.Results CCK-8 showed the proliferation inhibition rate of LOVO cells was increased gradually in oridonin-treated groups in a concentration and time dependent manner(all P<0.05).RT-PCR showed that the gene expression of the E-cadherin was increased and the gene expression of Vimentin was reduced in LOVO cells treated with 4,8 and 12μg/mL oridonin for 48 h(all P<0.05).The results of Western blot showed that the expression level of the p-GSK-3β(Tyr216)protein was up-regulated and the expression level of the p-GSK-3β(Ser9)andβ-catenin protein were down-regulated in LOVO cells treated with 4,8 and 12μg/mL oridonin for 48 h(all P<0.05).One-hour pretreatment of 2μmol/L GSK-3βspecific inhibitor CHIR and 24 h treatment of 2μmol/L CHIR and 12μg/mL oridonin could reverse the effect of oridonin on the expression ofβ-catenin,E-cadherin and Vimentin(all P<0.05).Conclusion Oridonin can inhibit the proliferation and EMT of colon cancer LOVO cells by Wnt/β-catenin signaling pathway.

关 键 词：结肠肿瘤 冬凌草甲素 WNT/Β-CATENIN信号通路 LOVO细胞 p-GSK-3β 

分 类 号：R735.35[医药卫生—肿瘤]