miR-34a-5p提高顺铂在宫颈癌细胞中的敏感性及其可能机制

miR-34a-5p enhances the sensitivity of cisplatin in cervical cancer cells and its possible mechanism

目的:探究miR-34a-5p对宫颈癌细胞顺铂敏感性的影响及其可能机制。方法:分别使用0、1、2、4、8μg/mL浓度的顺铂处理Hela细胞,CCK8检测细胞活力,qRT-PCR检测miR-34a-5p表达。设计并合成miR-34a-5p mimics,将细胞分成Blank、NC、miR-34a-5p、AG490、miR-34a-5p+AG490组。CCK8检测细胞活力,流式细胞术检测细胞凋亡率,Transwell实验检测侵袭细胞数,Western blot检测MDM4、STAT3、MRP1、Bax和BCL-2蛋白表达水平。结果:随着顺铂浓度的增加,Hela细胞活力和miR-34a-5p表达逐渐下降。与Blank/NC组相比,miR-34a-5p和AG490组细胞活力、侵袭细胞数、MRP1、BCL-2、MDM4、p-STAT3/STAT3蛋白表达显著下降(P<0.05),细胞凋亡率、Bax蛋白表达显著上升(P<0.05);与miR-34a-5p/AG490组相比,miR-34a-5p+AG490组细胞活力、侵袭细胞数、MRP1、BCL-2、MDM4、p-STAT3/STAT3蛋白表达显著下降(P<0.05),细胞凋亡率、Bax蛋白表达显著上升(P<0.05)。结论:miR-34a-5p可增加宫颈癌细胞对顺铂化疗的敏感性,其机制可能与MDM4/JAK/STAT3信号通路相关。

Objective:To explore the effect of miR-34a-5p on cisplatin sensitivity of cervical cancer cells and its possible mechanism.Methods:Hela cells were treated with cisplatin at concentrations of 0,1,2,4 and 8μg/mL respectively.Cell viability was detected by CCK8,and the expression of miR-34a-5p was detected by qRT-PCR.miR-34a-5p mimics were designed and synthesized,and the cells were divided into Blank,NC,miR-34a-5p,AG490 and miR-34a-5p+AG490 groups.Cell viability was detected by CCK8.Apoptosis rate was detected by flow cytometry.The number of invasive cells was detected by Transwell assay.The protein expression levels of MDM4,STAT3,MRP1,Bax and BCL-2 were detected by Western blot.Results:With the increase of cisplatin concentration,Hela cell viability and the expression of miR-34a-5p decreased gradually.After grouping,compared with the Blank/NC group,the cell viability,number of invasive cells,and the protein expression levels of MRP1,BCL-2,MDM4,p-STAT3/STAT3 in the miR-34a-5p and AG490 groups were decreased significantly(P<0.05),while the apoptosis rate and Bax protein expression were increased significantly(P<0.05).Compared with the miR-34a-5p/AG490 group,the cell viability,number of invasive cells,and the protein expression levels of MRP1,BCL-2,MDM4,p-STAT3/STAT3 in the miR-34a-5p+AG490 groups were decreased significantly(P<0.05),while the apoptosis rate and Bax protein expression were increased significantly(P<0.05).Conclusion:miR-34a-5p can increase the sensitivity of cervical cancer cells to cisplatin chemotherapy,and its mechanism may be related to MDM4/JAK/STAT3 signaling pathway.