MicroRNA Expression Profile Separates Squamous Cell Carcinoma by Mode of Differentiation.

Andani Marumo, Adam Botha, Julitha Molepo, Henry Adeola, Pumza Samantha Maganagane, Mulalo Molaudzi
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Abstract

Background: Squamous cell carcinoma (SCC) is a non-melanoma skin cancer with several risk factors including age and sun exposure. The degree of histological differentiation is considered an independent predictor of recurrence, metastasis, and survival. MicroRNAs (miRNAs) are small non-coding RNAs that play an important role in regulating gene expression, culminating in the initiation and progression of multiple tumors. The aim of this study was to determine changes in miRNA expression as a result of the mode of differentiation in SCC.

Methods: We analyzed 29 SCC samples that were separated by mode of differentiation into well (n=4), moderate (n=20) and poor (n=5). Of the 29 samples, five had matched normal tissues, which were used as controls. Total RNA was extracted using the RNeasy FFPE kit, and miRNAs were quantified using Qiagen MiRCURY LNA miRNA PCR Assays. Ten miRNAs (hsa-miR-21, hsa-miR-146b-3p, hsa-miR-155-5p, hsa-miR-451a, hsa-miR-196-5p, hsa-miR-221-5p, hsa-miR-375, hsa-miR-205-5p, hsa-let-7d-5p and hsa-miR-491-5p) that have been previously differentiated in cancer, were quantified. A fold regulation above 1 indicated upregulation and below 1, downregulation.

Results: Hierarchical clustering showed that the miRNA expression profile in the moderately differentiated group was similar to the well-differentiated group. The miRNA with the greatest upregulation in the moderate group was hsa-miR-375, while in the well group, hsa-miR-491-5p showed the greatest downregulation.

Conclusion: In conclusion, this study observed that the well and moderate groups had similar microRNA expression patterns compared to the poorly differentiated group. MicroRNA expression profiling may be used to better understand the factors underpinning mode of differentiation in SCC.

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微小RNA表达谱通过分化模式分离鳞状细胞癌。
背景:鳞状细胞癌(SCC)是一种癌症,有多种危险因素,包括年龄和日晒。组织学分化程度被认为是复发、转移和生存的独立预测因素。微小RNA(miRNA)是一种小型非编码RNA,在调节基因表达中发挥重要作用,最终导致多发性肿瘤的发生和发展。本研究的目的是确定SCC分化模式导致的miRNA表达的变化。方法:我们分析了29个SCC样本,这些样本按分化模式分为良(n=4)、中(n=20)和差(n=5)。在29个样本中,有5个样本与正常组织匹配,作为对照。使用RNeasy FFPE试剂盒提取总RNA,并使用Qiagen MiRCURY LNA miRNA PCR分析对miRNA进行定量。对先前在癌症中分化的10种miRNA(hsa-miR-21、hsa-miR-146b-3p、hsa-iR-155-5p、hsa-miR-451a、hsa--miR-196-5p、hsa-miR-221-5p、hsa-1miR-375、hsa-miR-205-5p、hsa-let-7d-5p和hsa-miR-491-5p)进行定量。倍数调节高于1表示上调,低于1表示下调。结果:分层聚类显示,中分化组和高分化组的miRNA表达谱相似。中度组中上调最大的miRNA是hsa-miR-375,而在正常组中,hsa-miR-491-5p表现出最大的下调。结论:总之,本研究观察到,与低分化组相比,高分化组和中分化组具有相似的微小RNA表达模式。微RNA表达谱可用于更好地了解SCC分化模式的基础因素。
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