miRNA-200c对非小细胞肺癌A549细胞甲氨蝶呤耐药性的影响  

Impact of miRNA-200c on Methotrexate Resistance of Non-small Cell Lung Cancer Cells A549

作　　者：单武林 邓芳[1] 张晓蕾[1] 张婧[1] 韩丹丹[1] 万玲玲[1] 李明[1] 

机构地区：[1]安徽省肿瘤医院(安徽省立医院西区)检验科,合肥230031

出　　处：《肿瘤防治研究》2016年第5期321-325,共5页Cancer Research on Prevention and Treatment

基　　金：安徽省自然科学基金(1308085QH144)

摘　　要：目的探讨miRNA-200c(miR-200c)在非小细胞肺癌A549细胞耐甲氨蝶呤(MTX)(A549/MTX)中的影响及可能的作用机制。方法通过实时荧光定量(q RT-PCR)法检测人肺癌亲本细胞株A549细胞、转染miR-200c模拟物(mimic)的A549/MTX细胞(A549/MTX-M)及转染miR-阴性对照(NC)A549/MTX细胞(A549/MTX-N)中miR-200c的表达水平。分别采用MTT法、锥虫兰染色及流式细胞术检测三组细胞对MTX的药物敏感度、细胞增殖能力及细胞凋亡变化,并采用q RT-PCR检测其P53和P21基因表达的变化。结果 miR-200c在A549细胞中的表达水平显著高于A549/MTX-N细胞;A549/MTX-M细胞miR-200c水平高于A549/MTX-N细胞;用不同浓度MTX刺激细胞,与A549/MTX-N细胞比较,A549/MTX-M细胞的增殖能力减弱、凋亡细胞增多,并呈剂量依赖性,差异均有统计学意义。转染miR-200c mimic后,P53和P21基因表达水平上升,与转染miR-NC细胞比较,差异有统计学意义。结论 miR-200c能够逆转A549/MTX细胞对MTX的耐药性,其作用机制可能是通过P53/P21信号转导通路诱导细胞凋亡来实现的。Objective To explore the effect of micro RNA-200c（miR-200c） on methotrexate（MTX） resistance of non-small cell lung cancer cells A549（A549/MTX） and elucidate its related mechanism. Methods Quantitative real-time PCR（q RT-PCR） was used to detect the miR-200 c expression in A549 cells, A549/MTX cells transfected with miR-200 c mimic（A549/MTX-M） and A549/MTX cells transfected with miR-negative control（A549/MTX-N）. MTT assay, Trypan blue staining and flow cytometry analysis were sequentially performed to detect the sensitivity of A549, A549/MTX-M and A549/MTX-N cells to MTX, cell proliferation and apoptosis. q RT-PCR was used to detect the gene expressions of P53 and P21 in these cells. Results The miR-200 c expression in A549 cells was significantly higher than that in A549/MTX-N cells. The miR-200 c expression in A549/MTX-M cells was significantly higher than that in A549/MTX-N cells. Compared with A549/MTX-N cells, the proliferation inhibition and apoptosis of A549/MTX-M cells were increased after treated with MTX in a concentration-dependent manner, with significant difference. Furthermore, the up-regulated P53 and P21 expression were observed in A549/MTX-M cells, with significant difference compared with A549/MTX-N cells（P=0.023, P=0.015）. Conclusion miR-200 c could reduce the resistance of A549/MTX cells to MTX with the possible mechanism of inducing the apoptosis through the P53/P21 pathway.

关 键 词：MIR-200C 甲氨蝶呤 耐药 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]