摘要
目的探讨桔梗皂苷-D(PD)联合5-氟尿嘧啶(5-FU)对宫颈癌细胞的增殖抑制作用。方法用CCK-8实验检测PD单用及与5-FU联用对宫颈癌细胞的增殖抑制作用及半抑制浓度(IC 50)。将细胞分为对照组、20μmol/L PD组、20μmol/L 5-FU组及联合处理组(10μmol/L PD+10μmol/L 5-FU)。用流式细胞术检测各组细胞凋亡率、线粒体膜电位变化和活性氧水平变化;用细胞划痕实验检测细胞迁移;用Western blot检测各组蛋白表达变化。结果CCK-8实验结果显示,联合处理组宫颈癌细胞的存活率降低,与两个单药实验组相比,差异有统计学意义(P<0.05)。流式细胞术显示联合处理组凋亡细胞比例达到42.37%,高于两个单药实验组,差异有统计学意义(P<0.05)。联合处理组迁移的细胞数目显著降低,抑制效果达到50%,与两个单药实验组相比差异有统计学意义(P<0.05)。流式细胞术显示联合处理组活性氧水平大幅度上升,加入NAC后联合处理组凋亡细胞比例下降为15.26%(P<0.05)。Western blot结果表明联合处理组葡萄糖转运蛋白-1(GLUT1)蛋白的表达量降低,加入NAC后GLUT1蛋白的表达升高。结论PD与5-FU联用能协同发挥抗癌作用,促进宫颈癌细胞凋亡,其作用机制是通过上调细胞中ROS水平,进而抑制GLUT1蛋白的表达。
Objective To explore the inhibitory effects of Platycodin-D(PD)combined with 5-fluorouracil(5-FU)on the proliferation of cervical cancer cells.Methods The inhibitory effect of PD alone and combined with 5-FU on the proliferation of cervical cancer cells and the IC 50 value were detected by CCK-8 assay.Subsequently,the cells were divided into the blank group,the 20μmol/L PD group,the 20μmol/L 5-FU group and the combined treatment group(10μmol/L PD+10μmol/L 5-FU),and the cells were treated with corresponding concentrations.The flow cytometry was used to detect cell apoptosis rate,mitochondrial membrane potential change and ROS level change in each group.Cell scratch test was used to detect cell migration.Western blot was used to detect changes of the protein expression in each group.Results The CCK-8 experiment results showed that the survival rate of the four cervical cancers in the combined treatment group was further reduced.Compared with the two single-agent experimental groups,the difference was significant(P<0.05).Flow cytometry showed that the proportion of apoptotic cells in the combined treatment group reached 42.37%,which was higher than the apoptosis-inducing effect of the two drugs when treated with 20μmol/L,and there was a significant difference(P<0.05).The combined treatment group significantly reduced the number of migrating cells,and the inhibitory effect reached 50%,the difference was statistically significant compared with the two drug groups alone(P<0.05).Flow cytometry showed that the level of reactive oxygen species in the combined treatment group increased significantly.After adding NAC,the percentage of apoptotic cells in the combined treatment group dropped significantly to 15.26%,which was significantly different from the combined treatment group(P<0.05).Western blot results showed that the expression of GLUT1 protein decreased after PD and 5-FU were combined,and the expression of GLUT1 protein increased after adding NAC.Conclusion The combination of PD and 5-FU can synergistically exert anti-cancer effects and greatly promote the apoptosis of cervical cancer cells.The mechanism is related to up-regulating the level of ROS in cells,thereby inhibiting the expression of GLUT1 protein.
作者
王立娟
孙全
曾敏
WANG Lijuan;SUN Quan;ZENG Min(Department of Pathology,Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400021,China;Department of Pharmacy,Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400021,China)
出处
《重庆医学》
CAS
2023年第12期1783-1788,共6页
Chongqing medicine
基金
重庆市科研机构绩效激励引导专项基金资助项目(cstc2019jxjl130009)。
