摘要
目的研究微小RNA-152(miRNA-152)在非小细胞肺癌(NSCLC)顺铂(DDP)耐药过程中的作用机制。方法实时荧光定量逆转录PCR(qRT-PCR)检测NSCLC细胞株A549及其DDP耐药株A549/DDP细胞内的miRNA-152水平。通过转染miRNA-52模拟物(miRNA-152mimic)以提高A549/DDP细胞内的microRNA-152水平。MTT试验、倒置相差显微镜技仪和流式细胞术观察上调miRNA-152对细胞增殖及凋亡的影响,同时采用qRT-PCR和Western blot技术观察细胞内Bcl-2及核转录因子-κB(NF-κB)水平变化。结果 A549/DDP细胞中存在miRNA-152的低表达,Bcl-2及NF-κB的高表达。上调miRNA-152后,DDP造成的A549/DDP细胞增殖抑制率和凋亡率显著高于未上调miRNA-152的细胞,差异有统计学意义(P<0.05)。此外,miRNA-152mimic转染可显著降低A549/DDP细胞中的Bcl-2及NF-κB表达。结论低miRNA-152表达可能引起NSCLC对DDP的耐药,miRNA-152可能通过调节Bcl-2及NF-κB的水平介导NSCLC细胞对顺铂的敏感性。
Objective To investigate the action mechanism of microRNA-152（miRNA-152）in the cisplatin（DDP）resistance process in non-small cell lung cancer（NSCLC）.Methods The miRNA-152 level in NSCLC cell line A549 and its cisplatin-resistant cell line A549/DDP was detected by the real time quantitative PCR（qRT-PCR）.miRNA-152 mimic was transfected for increasing the intracellular miRNA-152 level in A549/DDP.The MTT assay,inverted miroscope technique and flow cytometry were adopted to observe the effect of up-regulating miRNA-152 on cell proliferation inhibition and apoptosis,meanwhile,the level changes of intracellular Bcl-2and NF-κB were also observed by adopting qRT-PCR and Western blot.Results The low expression of miRNA-152 and the high expression of Bcl-2and NF-κB were found in A549/DDP cells.Up-regulation of miRNA-152 enhanced the inhibitory effect of DDP in A549/DDP cells.Furthermore,after up-regulating miRNA-152,the inhibiting rate and apoptosis rate of A549/DDP cells caused by DDP were significantly higher than those in the cells without up-regulating miRNA-152,the difference was statistically significant（P〈0.05）.In addition,miRNA-152 mimic transfection significantly decreased the expression of Bcl-2and NF-κB in A549/DDP cells.Conclusion Low expression of miRNA-152 may induce the resistance of NSCLC to DDP,miRNA-152 could mediate the sensitivity of NSCLC cells to DDP via regulating Bcl-2and NF-κB levels.
出处
《重庆医学》
CAS
北大核心
2016年第10期1297-1301,共5页
Chongqing medicine
