大车前苷抑制卵巢癌SKOV3细胞侵袭转移的作用研究

Research on Invasion and Metastasis of Plantamajoside Inhibits Ovarian Cancer SKOV3 Cells

目的 探讨大车前苷(plantamajoside,PMS)对卵巢癌SKOV3细胞侵袭转移的抑制作用及其机制。方法 噻唑蓝(methyl thiazolyl tetrazdium,MTT)法检测PMS对卵巢癌SKOV3细胞的增殖抑制作用;将卵巢癌SKOV3细胞分为对照组,模型组,低、中、高剂量组;除对照组外,其余各组用转化生长因子(transforming growth factor,TGF)-β1诱导产生上皮间质转化(epithelial mesenchymal transition,EMT);低、中、高剂量组使用不同浓度的PMS(20、40、80μmol/L)干预;划痕及Transwell迁移实验测定细胞迁移能力;Transwell侵袭实验测定细胞侵袭能力;蛋白免疫印迹(western blot)和实时荧光定量聚合酶链式反应(real time quantitative polymerase chain reaction, RT-qPCR)法检测E-钙黏蛋白(E-cadherin,E-ca)、N-钙黏蛋白(N-cadherin,N-ca)、波形蛋白(vimentin)、转录因子(snail)及m RNA表达情况。结果 PMS可以明显抑制卵巢癌SKOV3细胞的增殖,且具有时间、剂量依赖性;除对照组外,其余各组细胞均产生EMT;与模型组相比,低、中、高剂量组细胞相对迁移率、迁移及侵袭细胞数均显著降低,E-Ca蛋白及m RNA表达量显著升高(P<0.05,P<0.01),N-Ca、Vimentin和Snail蛋白及m RNA表达量显著降低(P<0.05,P<0.01),且具有剂量依赖性。结论 PMS能够抑制卵巢癌SKOV3细胞的侵袭转移,其机制可能是通过逆转EMT的发生。

Objective To investigate the inhibitory effect of Plantamajoside(PMS)on invasion and metastasis of ovarian cancer SKOV-3 cells and its mechanism.Methods methyl thiazolyl tetrazdium(MTT)was used to detect the proliferation and inhibitory effect of PMS on ovarian cancer SKOV3 cells.Ovarian cancer SKOV3 cells were divided into control group,model group,low,medium and high dose groups.Transforming growth factor-β1(TGF-β1)was used to induce epithelial-mesenchymal transition(EMT)in other groups except the control group.Low,medium and high dose groups were treated with different concentrations of PMS(20,40,80μmol/L).Transwell invasion assay was used to determine the cell invasion ability.Western blot and real time quantitative polymerase chain reaction(RT-qPCR)analysis of E-cadherin(E-Ca),N-cadherin(N-Ca),Vimentin,Snail protein and mRNA expression.Results PMS can significantly inhibit the proliferation of ovarian cancer SKOV3 cells,and has time,dose dependent laziness.Except the control group,cells in other groups produced EMT;compared with model group,low,medium and high dose groups,the number of migrating cells and invading cells were significantly decreased,the expression levels of E-Ca protein and mRNA were significantly increased(P<0.05 or P<0.01),N-Ca,Vimentin and Snail protein and mRNA were significantly decreased(P<0.05 or P<0.01),and has dose dependent laziness.Conclusion PMS can inhibit the invasion and metastasis of ovarian cancer SKOV3 cells,possibly by reversing the occurrence of EMT.