芳姜黄酮对人皮肤鳞状细胞癌A431细胞迁移、侵袭及凋亡的影响和机制研究

为研究芳姜黄酮(Ar-Turmerone)对人鳞状细胞癌A431细胞增殖、迁移、侵袭和凋亡的影响及机制。实验采用CCK-8法检测抑制率,吉姆萨染色观察细胞形态,划痕实验和Transwell小室实验研究细胞迁移和侵袭能力的变化,流式细胞仪检测细胞凋亡率。此外,通过实时荧光定量聚合酶链反应(Real-time PCR)与蛋白质印迹法(western blot)法检测mRNA和蛋白表达。siRNA阻断Notch1,Hes1和PTEN,检测相应的下游mRNA和蛋白的表达变化,流式细胞仪检测细胞凋亡率。结果发现,芳姜黄酮可以抑制A431细胞增殖,使细胞形态发生改变,抑制细胞体外迁移和侵袭能力,促进细胞凋亡。经过芳姜黄酮处理后,Notch1,Hes1,PTEN的mRNA和蛋白表达升高。沉默Notch1,Hes1 mRNA和蛋白表达低于单纯给药组,而沉默Hes1,PTEN mRNA和蛋白表达也低于单纯给药组;沉默PTEN后,与单纯给药组相比,细胞死亡率降低。总之,芳姜黄酮可以抑制人鳞状细胞癌A431细胞的增殖并促进其凋亡,且具有抑制体外迁移和侵袭的作用,其促进细胞凋亡的机制是通过Notch1/Hes1/PTEN途径实现的。

Effects of Ar-Turmerone on the migration,invasion and apoptosis of human squamous cell carcinoma A431 cells

In order to study the effects of Ar-Turmerone on proliferation,migration,invasion and apoptosis of human squamous cell carcinoma A431 cells,CCK-8 method was used to detect the inhibition rate of Ar-Turmerone on A431 cells.Cell morphology was observed by Giemsa staining,cell migration ability was analyzed by scratch assay;cell invasion ability was studied by Transwell chamber experiment,apoptosis rate was detected by flow cytometry.The expression of mRNA was detected by real-time PCR,and the protein was detected by western blotting.The expression of Notch1,Hes1 and PTEN was blocked by siRNA,the expression of corresponding downstream mRNA and protein was detected,and the apoptosis rate was detected by flow cytometry.It was found that the action of Ar-Turmerone on A431 cells can inhibit cell proliferation,affect cell morphology,and inhibit cell migration and invasion,and promote cell apoptosis.After treatment,Notch1,Hes1,PTEN mRNA and protein expression were elevated.Silencing Notch1,Hes1 mRNA and protein expression were lower than the simple administration group,while silencing Hes1,PTEN mRNA and protein expression was also lower than the simple administration group;after silencing PTEN,compared with the simple drug-administered group,the death rate of the cell is reduced.In conclusion,it can be proved that the Ar-Turmerone can inhibit the proliferation of human squamous cell carcinoma A431 cells and promote its apoptosis;it can inhibit migration and invasion in vitro;the mechanism of Ar-Turmerone promoting apoptosis is through Notch1/Hes1/PTEN pathway is implemented.