Tafsiri Elham, Darbouy Mojtaba, Shadmehr Mohammad Behgam, Cho William C, Karimipoor Morteza
Department of Molecular Medicine, Biotechnology Research Center, Pasture Institute of Iran, Tehran, Iran.
J Cancer Res Ther. 2016 Jan-Mar;12(1):395-400. doi: 10.4103/0973-1482.148673.
Lung cancer is one of the most serious types of cancer that often diagnosed at advanced stage. MicroRNAs (miRNAs) are small non-coding molecules which silence gene expression of target gene (s) at posttranscriptional level. They are key regulators of cell cycle, apoptosis, anti-cancer drug responsiveness and metastasis.
Identification of the differential expression level of miR-15a/16, miR-21, miR-34a, miR-126, miR-128 and miR-210 in A549 cell line versus normal tissues and their correlation with selected corresponding target genes.
A549 cell line was cultured in F-12K medium and miRNA was extracted from normal tissues (2-3 cm adjacent to tumor tissue) and A549 cell line. cDNA was synthesized with specific stem-loop primers for each miRNA, while OligodT primer was used for target genes cDNA synthesis. Real-time quantitative polymerase chain reaction. (RT-qPCR) was used to analyze the expression pattern of miRNAs and target genes in A549 and normal non-small cell lung carcinoma. (NSCLC) tissues.
miR-15a/16, miR-34a, miR-126 and miR-128 were down-regulated significantly. (>2-fold change), while miR-21 and miR.210 were up-regulated in A549. Bcl-2 as miR-34a target gene was down-regulated while Hif-1α and Akt-3 were up-regulated that might be miR-210 and miR-34a target genes, respectively.
The significant differential expression level of these miRNAs made them as candidate biomarkers in NSCLC tumor tissues of patients. Perhaps Bcl-2 down-regulation and Akt-3 up-regulation can be linked with survival signals in A549 cell line. We can conclude that Bcl-2 and Akt-3 might be therapeutic targets to inhibit cell proliferation in NSCLC.
肺癌是最严重的癌症类型之一,通常在晚期才被诊断出来。微小RNA(miRNA)是一类小的非编码分子,可在转录后水平使靶基因的基因表达沉默。它们是细胞周期、细胞凋亡、抗癌药物反应性和转移的关键调节因子。
鉴定A549细胞系与正常组织中miR-15a/16、miR-21、miR-34a、miR-126、miR-128和miR-210的差异表达水平及其与选定的相应靶基因的相关性。
将A549细胞系培养于F-12K培养基中,从正常组织(肿瘤组织相邻2-3厘米处)和A549细胞系中提取miRNA。使用针对每种miRNA的特异性茎环引物合成cDNA,而使用oligodT引物合成靶基因cDNA。采用实时定量聚合酶链反应(RT-qPCR)分析A549和正常非小细胞肺癌(NSCLC)组织中miRNA和靶基因的表达模式。
miR-15a/16、miR-34a、miR-126和miR-128显著下调(变化>2倍),而miR-21和miR-210在A549中上调。作为miR-34a靶基因的Bcl-下降,而Hif-1α和Akt-3上调,它们可能分别是miR-210和miR-34a的靶基因。
这些miRNA的显著差异表达水平使其成为NSCLC肿瘤组织中患者的候选生物标志物。也许Bcl-2下调和Akt-3上调可能与A549细胞系中的生存信号有关。我们可以得出结论,Bcl-2和Akt-3可能是抑制NSCLC细胞增殖的治疗靶点。