MicroRNA-196a promotes non-small cell lung cancer cell proliferation and invasion through targeting HOXA5.
MicroRNA-196a promotes non-small cell lung cancer cell proliferation and invasion through targeting HOXA5.
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MicroRNA-196a通过靶向HOXA5促进非小细胞肺癌细胞增殖和侵袭
DOI：
10.1186/1471-2407-12-348
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发表时间：
2012-08-09
期刊：
影响因子：
3.8
通讯作者：
Wang ZX
中科院分区：
文献类型：
--
作者：
Liu XH;Lu KH;Wang KM;Sun M;Zhang EB;Yang JS;Yin DD;Liu ZL;Zhou J;Liu ZJ;De W;Wang ZX
MicroRNAs(MiRNAs)是一种短小的、非编码的RNAs(~22nt)，通过负向调节基因表达，在人类疾病的发病机制中发挥重要作用。虽然miR-196A与其他几种癌症有关，但它在非小细胞肺癌(NSCLC)中的作用尚不清楚。本研究旨在探讨miR-196A在非小细胞肺癌组织中的表达模式及其临床意义，并探讨其在肿瘤进展中的生物学作用。方法采用定量逆转录聚合酶链式反应(qRT-PCR)方法检测34例NSCLC组织和5株NSCLC细胞系中miR-196A的表达。通过5-氮杂-2-脱氧胞苷处理和亚硫酸氢盐测序，研究DNA甲基化对miR-196A基因表达的影响。用四甲基偶氮唑盐比色法和集落形成实验检测miR-196A对细胞增殖的影响，用Transwell法检测细胞的迁移和侵袭能力。Western blotting分析靶蛋白的表达。构建了荧光素酶报告载体以证实miR-196A对包括HOXA5在内的下游靶基因的作用。结果iR-196A在非小细胞肺癌组织和细胞系中的表达均高于相应的正常对照， 去甲基化可能对其表达有影响。MiR-196A在NSCLC组织中的高表达与NSCLC的临床分期有关，也与NSCLC的淋巴结转移有关，体外功能分析表明miR-196A的表达调控影响NSCLC细胞的增殖、迁移和侵袭。我们的分析表明miR-196A在mRNA和蛋白水平上都抑制了HOXA5的表达，荧光素酶检测证实miR-196A直接与HOXA5的3‘非翻译区结合。RNAi下调A549细胞HOXA5的表达可促进NSCLC细胞的增殖、迁移和侵袭。结论miR-196A在非小细胞肺癌组织中的表达显著上调，并部分通过下调HOXA5的表达来调控细胞的增殖、迁移和侵袭。因此，miR-196A可能是非小细胞肺癌干预治疗的潜在靶点。
BackgroundMicroRNAs (miRNAs) are short, non-coding RNAs (~22 nt) that play important roles in the pathogenesis of human diseases by negatively regulating gene expression. Although miR-196a has been implicated in several other cancers, its role in non-small cell lung cancer (NSCLC) is unknown. The aim of the present study was to examine the expression pattern of miR-196a in NSCLC and its clinical significance, as well as its biological role in tumor progression.MethodsExpression of miR-196a was analyzed in 34 NSCLC tissues and five NSCLC cell lines by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The effect of DNA methylation on miR-196a expression was investigated by 5-aza-2-deoxy-cytidine treatment and bisulfite sequencing. The effect of miR-196a on proliferation was evaluated by MTT and colony formation assays, and cell migration and invasion were evaluated by transwell assays. Analysis of target protein expression was determined by western blotting. Luciferase reporter plasmids were constructed to confirm the action of miR-196a on downstream target genes, includingHOXA5. Differences between the results were tested for significance using Student’s t-test (two-tailed).ResultsmiR-196a was highly expressed both in NSCLC samples and cell lines compared with their corresponding normal counterparts, and the expression of miR-196a may be affected by DNA demethylation. Higher expression of miR-196a in NSCLC tissues was associated with a higher clinical stage, and also correlated with NSCLC lymph-node metastasis.In vitrofunctional assays demonstrated that modulation of miR-196a expression affected NSCLC cell proliferation, migration and invasion. Our analysis showed that miR-196a suppressed the expression of HOXA5 both at the mRNA and protein levels, and luciferase assays confirmed that miR-196a directly bound to the 3’untranslated region of HOXA5. Knockdown of HOXA5 expression in A549 cells using RNAi was shown to promote NSCLC cell proliferation, migration and invasion. Finally, we observed an inverse correlation between HOXA5 and miR-196a expression in NSCLC tissues.ConclusionsOur findings indicate that miR-196a is significantly up-regulated in NSCLC tissues, and regulates NSCLC cell proliferation, migration and invasion, partially via the down-regulation of HOXA5. Thus, miR-196a may represent a potential therapeutic target for NSCLC intervention.
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影响因子：
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作者：
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