Keloid microRNA expression analysis and the influence of miR-199a-5p on the proliferation of keloid fibroblasts
Keloid microRNA expression analysis and the influence of miR-199a-5p on the proliferation of keloid fibroblasts
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DOI：
10.4238/2014.april.14.2
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发表时间：
2014-01-01
影响因子：
0.4
通讯作者：
Luo, S-J
中科院分区：
文献类型：
--
作者：
Wu, Z-Y;Lu, L.;Luo, S-J
本研究的目的是鉴定参与瘢痕疙瘩形成的microRNA（miRNAs），并确定它们对瘢痕疙瘩成纤维细胞（KFs）增殖的影响。收集切除的瘢痕疙瘩组织和正常皮肤组织各8个标本。使用miRNA微阵列检测瘢痕疙瘩组织和正常皮肤中差异表达的miRNA，并通过定量实时聚合酶链反应（RT-PCR）进行验证。通过微阵列杂交鉴定了17个差异表达的miRNAs，包括miR-199 a-5 p。qRT-PCR分析证实了微阵列分析检测到的瘢痕疙瘩中miR-199 a-5 p表达相对于正常组织降低。然后将差异表达的miRNA的模拟物转染到KF细胞系中，并使用EdU测定法测定miRNA过表达对KF增殖的影响。与模拟转染的细胞相比，用miR-199 a-5 p模拟物转染的KFs显示出显著较低的细胞增殖和改变的细胞周期，细胞具有显著较长的S和G2/M期。瘢痕疙瘩组织中miRNA-199 a-5 p的低表达可能影响了KF的细胞周期，抑制了KF的增殖，提示miR-199 a-5 p可能在KF的增殖调控中发挥作用。
The purpose of this study was to identify microRNAs (miRNAs) involved in keloid formation and determine their influence on the proliferation of keloid fibroblasts (KFs). Eight specimens each of resected keloid tissue and normal skin tissue were collected. miRNAs that are differentially expressed in keloid tissue and normal skin were detected using an miRNA microarray and verified by quantitative real-time polymerase chain reaction (RT-PCR). Seventeen differentially expressed miRNAs, including miR-199a-5p, were identified by microarray hybridization. qRT-PCR analysis confirmed the decrease in miR-199a-5p expression in keloid vs normal tissue that was detected by the microarray analysis. Mimics of differentially expressed miRNAs were then transfected into a KF cell line, and the effect of miRNA overexpression on the proliferation of KFs was assayed using the EdU assay. Compared with mock-transfected cells, KFs transfected with a miR-199a-5p mimic showed significantly lower cell proliferation and an altered cell cycle, with cells having significantly longer S and G2/M phases. The significantly lower expression of miRNA-199a-5p in keloids likely influences the cell cycle of KFs and restrains their proliferation, suggesting that miR-199a-5p probably plays a role in the regulation of KF proliferation.