miR-223-3P对肺鳞癌细胞顺铂敏感性的影响

The role of miR-223-3p on drug resistance to cisplatin in lung squamous cell carcinoma

目的研究miR-223-3p对肺鳞癌细胞顺铂(CDDP)敏感性的影响。方法通过分别升高或降低(miR-223-3p mimics、NC-mimics、miR-223-3p inhibitor、NC-inhibitor)肺鳞癌NCI-H520和SK-MES-1细胞内miR-223-3p的含量后,qRTPCR检测细胞内miR-223-3p基因表达水平;CCK-8法检测细胞半数抑制浓度(IC50);流式细胞仪检测细胞加入CDDP药物刺激后凋亡率的改变(分别设计四组不同的处理:miR-223-3p mimics+CDDP、NC-mimics、miR-223-3p inhibitor+CDDP、NC-inhibitor);Transwell实验检测SK-MES-1细胞迁移能力的变化;Western blot法检测SK-MES-1细胞中Bax、ZEB1和E-cadherin的变化。结果CCK-8实验结果表明,miR-223-3p mimics组的IC50低于对照组的IC50,miR-223-3p inhibitor组的IC50高于对照组的IC50;流式细胞术凋亡检测结果显示,miR-223-3p mimics+CDDP(80μmol/L)组细胞凋亡率增加,而miR-223-3p inhibitor+CDDP(80μmol/L)组细胞凋亡率减少;Western blot证实,同时分别升高或降低肺鳞癌细胞内的miR-223-3p含量后,ZEB1蛋白相应的降低和升高,且Bax、E-cadherin蛋白相应的升高和降低。结论miR-223-3p作为抑癌基因影响肺鳞癌细胞对CDDP的敏感性,可作为治疗肺鳞癌CDDP耐药的重要分子靶标。

Objective To investigate the effect of miR-223-3p on cisplatin sensitivity of lung squamous cell carcinoma.Methods After increasing or decreasing(miR-223-3p mimics,NC-mimics,miR-223-3p inhibitor,NC-inhibitor)the content of miR-223-3p in lung squamous carcinoma NCI-H520 and SK-MES-1 cells,the changes of miR-223-3p in cells were detected by qRT-PCR,half inhibitory concentration(IC50)was detected by CCK-8 assay,apoptosis was detected by flow cytometry(four different treatments were designed:miR-223-3p mimics+CDDP,NC-mimics,miR-223-3p inhibitor+CDDP,NC-inhibitor),the migration ability of SK-MES-1 cells was detected by Transwell assay,the proteins of Bax,ZEB1 and E-cadherin in SK-MES-1 cells were detected by Western blot.Results The results of CCK-8 experiment showed that the IC50 of the miR-223-3p mimics group was lower than that of the control group,and the IC50 of the miR-223-3p group was higher than that of the control group.Flow cytometry apoptosis detection results showed that the apoptosis rate of miR-223-3p mimics+cisplatin(CDDP)group increased,while the apoptosis rate of miR-223-3p inhibitor+cisplatin(CDDP)group decreased.Western blot confirmed that ZEB1 protein,Bax and E-cadherin protein increased and decreased respectively after the content of miR-223-3p in lung squamous cell carcinoma was increased or decreased.Conclusion miR-223-3p affects the sensitivity of lung squamous cell carcinoma cells to cisplatin as an anticancer gene,and it can be an important molecular target for the treatment of cisplatin resistance in lung squamous cell carcinoma.