楤木皂苷C对乳腺癌MCF-7细胞增殖和迁移的影响研究

Effect of aralosdie C on proliferation and migration of breast cancer MCF-7 cells

目的研究楤木皂苷C对乳腺癌MCF-7细胞增殖、迁移和蛋白激酶B(Akt)通路活性的影响。方法将处于对数期的MCF-7细胞分为对照组(生理盐水处理)和低、中、高剂量实验组(4. 00,8. 00,20. 00μmo L·L-1楤木皂苷C处理)。以噻唑蓝(MTT)及Wound Healing法分别检测细胞增殖、迁移能力,以蛋白质印迹法检测细胞Akt、磷酸化Akt(p-Akt)、钙黏蛋白E(E-cadherin)和波形蛋白(Vimentin)的表达。结果干预24 h,低、中、高剂量实验组MCF-7细胞增殖抑制率分别为(10. 54±2. 28)%,(35. 43±2. 87)%,(60. 65±2. 14)%;干预48 h,分别为(14. 33±2. 13)%,(42. 57±2. 43)%,(72. 54±2. 27)%;干预72 h,分别为(22. 66±2. 37)%,(50. 54±2. 29)%,(85. 59±2. 49)%。干预24 h时,对照组和低、中、高剂量实验组划痕愈合率分别为(42. 13±3. 43)%,(31. 68±3. 63)%,(25. 13±3. 37)%,(17. 63±2. 57)%。蛋白质印迹显示,对照组及低、中、高剂量实验组Akt蛋白相对表达量分别为1. 23±0. 22,0. 85±0. 21,0. 72±0. 13,0. 57±0. 11,p-Akt蛋白表达量分别为0. 95±0. 09,0. 88±0. 09,0. 78±0. 11,0. 61±0. 12,E-cadherin蛋白相对表达量分别为0. 25±0. 06,0. 36±0. 04,0. 69±0. 11,1. 02±0. 15,Vimentin蛋白表达量分别为1. 26±0. 22,0. 86±0. 12,0. 53±0. 06,0. 39±0. 05;差异均有统计学意义(均P <0. 05)。结论楤木皂苷C可有效抑制乳腺癌MCF-7细胞增殖和迁移,其作用机制可能与抑制上皮细胞间充质化(EMT)及Akt通路活性相关。

Objective To investigate the inhibitory effect of aralosdie C on proliferation,migration and protein kinase B(Akt)pathway activity of breast cancer MCF-7 cells.Methods MCF-7 cells in logarithmic phase were divided into control group(normal saline)and low,middle,high dose experimental groups(4.00,8.00,20.00μmoL·L-1 aralosdie C).Cell proliferation and migration were detected by MTT and Wound Healing.The expression of Akt,phosphorylation-Akt(p-Akt),E-cadherin,Vimentin protein were detected by Western blot(WB).Results After 24 h of culture,the proliferation inhibition rate of MCF-7 cells in low,middle,high dose experimental groups were(10.54±2.28)%,(35.43±2.87)%,(60.65±2.14)%,after 48 h of culture,which were(14.33±2.13)%,(42.57±2.43)%,(72.54±2.27)%,after 72 h of culture,which were(22.66±2.37)%,(50.54±2.29)%,(85.59±2.49)%.After 24 h of culture,the scratch healing rates in control group and low,middle,high dose experimental groups were(42.13±3.43)%,(31.68±3.63)%,(25.13±3.37)%,(17.63±2.57)%,the relative expression of Akt protein were 1.23±0.22,0.85±0.21,0.72±0.13,0.57±0.11,and the relative expression of p-Akt protein were0.95±0.09,0.88±0.09,0.78±0.11,0.61±0.12,the relative expression of E-cadherin protein were0.25±0.06,0.36±0.04,0.69±0.11,1.02±0.15,the relative expression of Vimentin protein were 1.26±0.22,0.86±0.12,0.53±0.06,0.39±0.05,all with significant difference(all P<0.05).Conclusion Aralosdie C can effectively inhibit the proliferation and migration of breast cancer MCF-7 cells,and its mechanism may be related to the inhibition of the activities of epithelial mesenchymal(EMT)and Akt pathway activity.