MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh, EH4 2XU, UK.
Br J Cancer. 2012 Jan 31;106(3):553-61. doi: 10.1038/bjc.2011.568. Epub 2012 Jan 5.
The incidence of malignant melanoma is increasing faster than that for any other cancer. Histological examination of skin excision biopsies remains the standard method for melanoma diagnosis and prognosis. Significant morphological overlap between benign and malignant lesions complicates diagnosis, and tumour thickness is not always an accurate predictor of prognosis.
To identify improved molecular markers to support histological examination, we used microarray analysis of formalin-fixed and paraffin-embedded samples from different stages of melanomagenesis to identify differentially expressed microRNAs (miRNAs). Differential expression was validated by qRT-PCR, and functional studies were carried out after transfection of miRNA precursors or inhibitors into melanoma cells to modulate miRNA expression.
In all, 20 miRNAs showed highly significant differential expression between benign naevi and either primary or metastatic melanomas, the majority being downregulated in melanoma, whereas only 2 miRNAs, namely miR-203 and miR-205, were differentially expressed between primary and metastatic melanomas. In functional in vitro assays, overexpression of miR-200c and miR-205 inhibited anchorage-independent colony formation and overexpression of miR-211 inhibited both anchorage-independent colony formation and invasion.
We have identified a series of differentially expressed miRNAs that could be useful as diagnostic or prognostic markers for melanoma and have shown that three miRNAs (namely miR-200c, miR-205 and miR-211) act as tumour suppressors.
恶性黑色素瘤的发病率增长速度比其他任何癌症都要快。皮肤切除活检的组织学检查仍然是诊断和预测黑色素瘤预后的标准方法。良性和恶性病变之间存在显著的形态学重叠,这使得诊断变得复杂,而且肿瘤厚度并不总是准确的预后预测指标。
为了确定能够支持组织学检查的改良分子标志物,我们使用了微阵列分析方法,对黑色素瘤发生过程中不同阶段的福尔马林固定石蜡包埋样本进行分析,以鉴定差异表达的 microRNAs(miRNAs)。通过 qRT-PCR 对差异表达进行验证,并在转染 miRNA 前体或抑制剂以调节 miRNA 表达后进行功能研究。
共有 20 个 miRNAs 在良性痣与原发性或转移性黑色素瘤之间表现出高度显著的差异表达,大多数在黑色素瘤中下调,而只有 2 个 miRNAs,即 miR-203 和 miR-205,在原发性和转移性黑色素瘤之间表达差异。在体外功能测定中,miR-200c 和 miR-205 的过表达抑制了非锚定依赖性集落形成,而 miR-211 的过表达抑制了非锚定依赖性集落形成和侵袭。
我们已经确定了一系列差异表达的 miRNAs,它们可以作为黑色素瘤的诊断或预后标志物,并且已经表明三个 miRNAs(miR-200c、miR-205 和 miR-211)作为肿瘤抑制因子发挥作用。