紫草素对卵巢癌细胞SKOV3/DDP顺铂耐药的逆转作用

Reversal effect of shikonin on cisplatin resistance of ovarian cancer SKOV3/DDP cells

目的:探讨紫草素是否逆转卵巢癌细胞SKOV3/DDP的顺铂耐药效应及其作用机制。方法:采用CCK-8法确定紫草素和顺铂的最佳作用条件,流式细胞术检测细胞的周期分布及凋亡率;Western blot检测周期及凋亡相关调控因子细胞周期蛋白D1(cyclin D1)、周期蛋白依赖性激酶2(CDK2)、P18、p-Rb、Bcl-2、Bax和cleaved caspase-3的蛋白水平。结果:CCK-8实验结果显示,相较于单用顺铂,联合使用紫草素对顺铂耐药卵巢癌细胞SKOV3/DDP的生长抑制作用较为显著。此外,联用紫草素和顺铂可显著抑制细胞周期G_1/S转化,并增加细胞早期凋亡率。Western blot结果显示,相较于顺铂处理组,紫草素和顺铂联用组的cyclin D1、CDK2、p-Rb及Bcl-2的蛋白水平显著降低,而P18、Bax及cleaved caspase-3的蛋白表达显著增加。结论:紫草素可逆转卵巢癌SKOV3/DDP细胞的顺铂耐药效应,其作用机制可能与影响细胞周期及凋亡相关因子的表达,进而抑制细胞活力促进细胞凋亡相关。

AIM:To explore the reversal effect of shikonin on cisplatin resistance of ovarian cancer SKOV3/DDP cells and its potential mechanism.METHODS:The proper conditions of treatment with shikonin and cisplatin were determined by CCK-8 assay.The cell cycle and apoptotic rate were analyzed by flow cytometry.The protein levels of cell cycle-and apoptotic-related molecules,such as cyclin D1,cyclin-dependent kinases 2(CDK2),P18,p-Rb,Bcl-2,Bax and cleaved caspase-3,were determined by Western blot.RESULTS:The results of CCK-8 assay showed that compared with cisplatin group,combined treatment with shikonin and cisplatin had a better inhibitory effect on the growth of cisplatin-resistant SKOV3/DDP cells.The cell cycle G 1/S transition was inhibited,while early apoptotic rate was increased after combined use of shikonin and cisplatin.The results of Western blot showed that compared with cisplatin group,the cells in combination group had lower protein levels of cyclin D1,CDK2,p-Rb and Bcl-2,accompanied with higher protein levels of P18,Bax and cleaved caspase-3.CONCLUSION:Shikonin reverses the cisplatin resistance of ovarian cancer SKOV3/DDP cells.The mechanism may be related to the regulation of cell cycle-and apoptotic-related molecules,and further inhibition of cell viability and promotion of cell apoptosis.