和厚朴酚通过抑制Smad2/3磷酸化对子宫内膜癌细胞迁移及侵袭的调控作用

Regulatory Effect of Honokiol on Migration and Invasion of Endometrial Cancer Cells via Inhibiting Smad2/3 Phosphorylation

目的研究和厚朴酚通过抑制Smad2/3磷酸化对子宫内膜癌细胞迁移及侵袭的调控作用。方法培养子宫内膜癌RL95-2细胞株并分组给药,对照组用不含药物的培养基处理,不同剂量和厚朴酚组分别用含有5μg/ml、10μg/ml、15μg/ml和厚朴酚的培养基处理,15μg/ml和厚朴酚+20 ng/ml TGF-β1组用含有15μg/ml和厚朴酚和20 ng/ml TGF-β1的培养基处理。检测细胞迁移率,侵袭数目,迁移基因E-cadherin、N-cadherin、Vimentin及侵袭基因MMP2、MMP9、MMP14的mRNA表达水平,p-Smad2、p-Smad3的蛋白表达水平。结果5μg/ml、10μg/ml、15μg/ml和厚朴酚组RL95-2细胞的迁移率、侵袭数目、N-cadherin、Vimentin、MMP2、MMP9、MMP14的mRNA表达水平、p-Smad2、p-Smad3的蛋白表达水平均低于对照组,E-cadherin的表达水平均高于对照组,差异有统计学意义(P<0.05),且和厚朴酚剂量越高、上述变化越显著;15μg/ml和厚朴酚+20 ng/ml TGF-β1组RL95-2细胞的迁移率、侵袭数目、N-cadherin、Vimentin、MMP2、MMP9、MMP14的mRNA表达水平、p-Smad2、p-Smad3的蛋白表达水平均高于15μg/ml和厚朴酚组,E-cadherin的表达水平均低于15μg/ml和厚朴酚组,差异有统计学意义(P<0.05)。结论和厚朴酚抑制子宫内膜癌细胞的迁移和侵袭,该作用与抑制Smad2/3的磷酸化有关。

Objective To study the regulatory effect of honokiol on the migration and invasion of endometrial cancer cells via inhibiting Smad2/3 phosphorylation.Methods Endometrial cancer RL95-2 cell lines were cultured and divided into groups for drug administration.The control group was treated with culture medium without drugs,and the honokiol groups with different doses were treated with culture medium containing 5μg/ml,10μg/ml,and 15μg/ml honokiol.The 15μg/ml honokiol+20 ng/ml TGF-β1 group was treated with a culture medium containing 15μg/ml honokiol and 20 ng/ml TGF-β1.The cell migration rate,invasion number,mRNA expression levels of migration genes E-cadherin,N-cadherin,and Vimentin and invasion genes MMP2,MMP9,and MMP14,and the protein expression levels of p-Smad2 and p-smad3 were detected.Results In honokiol groups of 5μg/ml,10μg/ml,and 15μg/ml,the RL95-2 cell migration rate,invasion number,mRNA expression levels of N-cadherin,Vimentin,MMP2,MMP9,and MMP14,and protein expression levels of p-Smad2 and p-smad3 were lower than those in the control group,and the mRNA expression level of E-cadherin was higher than those in the control group,with the difference statistically significant(P<0.05).A higher dose of honokiol indicated more significant changes.The RL95-2 cell migration rate,invasion number,mRNA expression levels of N-cadherin,Vimentin,MMP2,MMP9,and MMP14,and protein expression levels of p-Smad2 and p-smad3 of the 15μg/ml honokiol+20 ng/ml TGF-β1 group were all higher than those of the 15μg/ml honokiol group,and the mRNA expression level of E-cadherin was lower than that of 15μg/ml honokiol group(P<0.05).Conclusion Honokiol inhibits the migration and invasion of endometrial cancer cells,which is related to the inhibition of Smad2/3 phosphorylation.