microRNA-3163靶向乳腺癌耐药蛋白逆转肺癌细胞多药耐药作用

miRNA-3163 Represses Breast Cancer Resistance Protein to Reverse Multi-Drug Resistance Process of Human Lung Cancer Cells

目的:验证microRNA-3163(miR-3163)在肺癌细胞中是否靶向乳腺癌耐药蛋白(BCRP),探索逆转肺癌细胞抗肿瘤药物多药耐药(MDR)的干预策略。方法:检测BCRP在肺癌细胞A549和耐药细胞系A549/ADR中的表达,利用系列浓度梯度的抗肿瘤药物处理细胞,计算其作用的IC50值;在A549/ADR细胞中转染miR-3163的模拟物或抑制剂,用Western印迹检测BCRP的表达水平;在此基础上检测miR-3163对抗肿瘤药物杀伤A549/ADR细胞的影响。结果:与A549细胞相比,A549/ADR细胞具有对抗肿瘤药物的MDR特性,BCRP在A549/ADR细胞中的表达显著上调。转染miR-3163的模拟物能够上调A549/ADR细胞对抗肿瘤药物阿霉素、紫杉醇、吉西他滨和吉非替尼的敏感性,逆转其MDR作用。这些抗肿瘤药物作用的IC50值分别从4.86±0.33、0.41±0.05、3.79±0.26和5.51±0.25μmol/L下调至0.30±0.05、0.07±0.01、0.67±0.10和1.58±0.42μmol/L。特异性实验结果表明,miR-3163的模拟物能够在A549/ADR细胞中下调BCRP的表达水平,转染miR-3163的抑制剂能够阻断miR-3163模拟物的作用。结论:miR-3163有可能通过靶向耐药蛋白BCRP逆转肺癌细胞的MDR作用。

Objective： To identify whether breast cancer resistance protein（BCRP） is a target of miR-3163 andthe role of miR-3163 in lung cancer cells multi-drug resistance（MDR） process. Methods： The MDR of A549 orA549/ADR cells was analyzed by exposure to anti-tumor drugs. The expression of BCRP in A549 or A549/ADRcells was identified by Western blotting analysis. A549/ADR cells transfected with miR-3163 mimics or its inhibi-tor were harvested for MTT assays. Inhibition rate and the IC50 value of anti-tumor drugs were calculated. Results： Compared with A549 cells, A549/ADR cells had a MDR feature with up-regulated BCRP. Transfection ofmiR-3163 mimics significantly enhanced the sensitivity of A549/ADR cells to anti-tumor drugs adriamycin, pacli-taxel, gemcitabine and gefitinib, indicated by IC50 value decreased from 4.86±0.33, 0.41±0.05, 3.79±0.26 or 5.51±0.25 μmol/L to 0.30±0.05, 0.07±0.01, 0.67±0.10 or 1.58±0.42 μmol/L, respectively. The miR-3163 mimics hadthe effect of repressing BCRP in A549/ADR cells, which was reversed by miR-3163 inhibitor. Conclusion： MDRprocess of lung cancer cells can be reversed by Mi R-3163 through BCRP repression.