不同浓度白头翁皂苷对口腔鳞癌细胞增殖、迁移和钙蛋白酶1、E-cadherin、N-cadherin蛋白的影响

Effects of Pulchinenoside at Different Concentrations on the Proliferation and Migration of Squamous Cell Carcinoma Cells and Calpain 1,E-cadherin,N-cadherin Protein

目的:探究白头翁皂苷对口腔鳞癌细胞增殖、迁移及钙蛋白酶1(calpain1)表达的影响。方法:用0,1.0,2.0,4.0,8.0,12.0 mg·L^(-1)白头翁皂苷处理人口腔鳞癌CAL27细胞。四甲基偶氮唑(MTT)法检测白头翁皂苷对CAL27细胞增殖的影响;细胞划痕实验与Transwell实验检测白头翁皂苷对CAL27细胞迁移与侵袭能力的影响;蛋白质印迹(Western blot)技术检测CAL27细胞calpain1、E-cadherin、N-cadherin蛋白的表达变化。结果:与空白对照组比较,白头翁皂苷对CAL27细胞的增殖抑制率显著增加(P<0.05),且具有时间-剂量依赖关系。12.0 mg·L^(-1)白头翁皂苷处理48 h后CAL27细胞细胞形态从长梭形间质样变成鹅卵石状上皮样。随着白头翁皂苷处理浓度的增加,CAL27细胞迁移与侵袭能力逐渐减弱(P<0.05);CAL27细胞中calpain1、N-cadherin蛋白表达水平逐渐降低(P<0.05),E-cadherin蛋白表达水平逐渐升高(P>0.05)。结论:白头翁皂苷可抑制口腔鳞癌CAL27细胞增殖、迁移与侵袭过程,其机制可能与calpain1、N-cadherin表达下调、E-cadherin表达上调有关。

Objective： To explore the influences of pulchinenoside on the proliferation and migration of oral squamous cell carcinoma cells and the expression of calpain 1（ calpain 1）. Methods： The human oral squamous cell carcinoma CAL27 cells were treated with pulchinenoside at the concentrations of 0,1. 0,2. 0,4. 0,8. 0,12. 0 mg·L-1. MTT method was used to detect the effect of pulchinenoside on the proliferation of CAL27 cells; the effects of pulchinenoside on the proliferation and migration of CAL27 cells were detected by cell scratch test and Transwell experiment; the expression changes of calpain 1,E-cadherin and N-cadherin protein in CAL27 cells were detected by Western blot. Results： MTT results showed that,compared with the control group,the inhibitory rate of pulchinenoside for the proliferation of CAL27 cells increased significantly（ P〈0. 05） in a time-and dose-dependent manner. The light microscope results showed that the CAL27 cells morphology changed from the long spindle shape to the cobblestone like epithelioid after the 48-hour treatment with 12. 0 mg·L-1 pulchinenoside. Cell scratch test results showed that with the increase of pulchinenoside concentration,the CAL27 cell migration ability gradually weakened. Transwell experimental results show that with the increase of pulchinenoside concentration,the CAL27 cell migration and invasion abilities decreased gradually（ P 〈 0. 05）. Western blot results showed that with the increase of the concentration of pulchinenoside,the expression levels of calpain 1 and N-cadherin protein in CAL27 cells decreased gradually（ P〈0. 05）,the expression level of E-cadherin protein increased gradually（ P〉0. 05）. Conclusion：Pulchinenoside can inhibit the proliferation,migration and invasion of oral squamous cell carcinoma CAL27 cells,and its mechanism may be related to the down-regulation of calpain 1 and N-cadherin expressions and up-regulation of E-cadherin expression.