MicroRNA-221对非小细胞肺癌NCI-H1299细胞生长的影响及其机制研究

Effect of microRNA-221 on NCI-H1299 Cell Growth in Non-small-cell Lung Cancer

目的研究microRNA-221(miRNA-221)对非小细胞肺癌NCI-H1299细胞株生长调节的影响。方法通过转染miR-221-mimics或miR-221-inhibitor建立miRNA-221过表达或下调的NCI-H1299细胞系;通过MTT试验和Brd U插入染色检测细胞活力和增殖能力的变化;使用Q-PCR和western blot检测增殖相关基因mRNA或蛋白表达的变化。结果 miRNA-221过表达能够增强NCI-H1299细胞的活力和增殖能力(P<0.01);P27是NCI-H1299细胞中miRNA-221的潜在靶基因,同时P27蛋白水平中也被miRNA-221负调控(P<0.01)。结论 miRNA-221可以促进非小细胞肺癌NCI-H1299细胞的生长,可作为非小细胞肺癌基因治疗的有效靶点。

Objective To investigate effect of microRNA-221（ miR-221） on growth regulation function of cell line NCI-H1299 in non-small-cell lung cancer（ NSCLC）. Methods NCI-H1299 cell lines of over-expressed miRNA-221 and down-regulated NCI-H1299 were established by transfecting miRNA-221 mimics or miR-221-inhibitor. Changes of cell viability and reproductive activity were detected using thiazolyl blue（ MTT） assay and Brd U immunostaining method. The mRNA and albumen expression changes of related proliferation genes were detected using quantitative-polymerase chain reaction（ Q-PCR） and western blot methods. Results Over-expressed miRNA-221 led to enhancement of cell viability and reproductive activity of NCI-H1299 cells（ P〈0. 01）. P27 was a potential target gene of miRNA-221 in NCIH1299 cell,and P27 was negatively regulated by miRNA-221 in the protein level（ P〈0. 01）. Conclusion MiR-221 can enhance the growth of cell line NCI-H1299 in non-small-cell lung cancer,and therefore it can be used as an effective target for gene therapy of non-small-cell lung cancer.