Dias S, Hemmings S, Muller C, Louw J, Pheiffer C
Biomedical Research and Innovation Platform (BRIP), South African Medical Research Council, Tygerberg, South Africa.
Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg, South Africa.
Biomed Res Int. 2017;2017:1080157. doi: 10.1155/2017/1080157. Epub 2017 Apr 26.
Dysregulated microRNA (miRNA) expression is observed during type 2 diabetes (T2D), although the consistency of miRNA expression across measurement platform and biological source is uncertain. Here we report miRNA profiling in the whole blood and serum of South African women with different levels of glucose tolerance, using next generation sequencing (NGS) and quantitative real time PCR (qRT-PCR). Whole blood-derived miRNAs from women with newly diagnosed T2D ( = 4), impaired glucose tolerance (IGT) ( = 4), and normal glucose tolerance (NGT) ( = 4) were subjected to NGS, whereafter transcript levels of selected miRNAs were quantified in the whole blood and serum of these women using qRT-PCR. Of the five significantly differentially expressed miRNAs identified by NGS, only the directional increase of miR-27b in women with IGT compared to NGT was confirmed in whole blood and serum, using qRT-PCR. Functional enrichment of miR-27b gene targets identified biological pathways associated with glucose transport and insulin regulation. In conclusion, this study showed poor correlation in miRNA expression profiled using NGS and qRT-PCR and in whole blood and serum. The consistent increased expression of miR-27b in women with IGT compared to NGT across measurement platform and biological source holds potential as a biomarker for risk stratification in our population.
在2型糖尿病(T2D)期间观察到微小RNA(miRNA)表达失调,尽管跨测量平台和生物来源的miRNA表达一致性尚不确定。在此,我们报告了使用下一代测序(NGS)和定量实时PCR(qRT-PCR)对不同糖耐量水平的南非女性全血和血清中的miRNA进行分析。对新诊断为T2D(n = 4)、糖耐量受损(IGT)(n = 4)和糖耐量正常(NGT)(n = 4)的女性全血来源的miRNA进行NGS检测,随后使用qRT-PCR对这些女性全血和血清中选定miRNA的转录水平进行定量。在NGS鉴定出的五个显著差异表达的miRNA中,使用qRT-PCR仅证实了IGT女性与NGT女性相比,miR-27b在全血和血清中的表达呈定向增加。miR-27b基因靶点的功能富集确定了与葡萄糖转运和胰岛素调节相关的生物学途径。总之,本研究表明,使用NGS和qRT-PCR以及在全血和血清中进行的miRNA表达谱分析相关性较差。与NGT女性相比,IGT女性中miR-27b在跨测量平台和生物来源中的表达持续增加,具有作为我们人群风险分层生物标志物的潜力。