The mechanisms of transcriptional regulation by microRMAs in relation to COLIA2 promotor region in keloid

本研究旨在研究瘢痕疙瘩组织中microRNAs(MiRNAs)的表达谱，以了解瘢痕疙瘩发病的分子机制(S)。我们证实miR-10a在瘢痕疙瘩成纤维细胞(KFS)中的表达明显低于正常真皮成纤维细胞(NFs)。用miRNA抑制剂和维甲酸(RA)分别抑制和诱导KFS和NFSmiR-10a的表达表明，MIR-10a抑制和诱导KFS培养上清液中IL-6和I型胶原原(PICP)的分泌，而RA处理KFS后，MIR-10a的分泌增加。这些结果提示miR-10a对KFS和NFS胶原的合成和分泌有调节作用。将miR-10a寡核苷酸导入KF和NF细胞。转染miR-10a的KFS和NFSPICP分泌减少。经RA处理后，miR-10a在KFS和NFS中高表达。收集处理后的细胞进行miR-10a定量聚合酶链式反应，并用酶联免疫吸附试验检测培养上清液中IL-6和PICP值。检测结果表明，RA治疗后KFS中IL-6浓度明显高于非KFS。我们的数据证实了miR-10a在KFS中的差异表达。MiR-10a调控KFS和NFS胶原的合成和分泌。MiR-10a表达下调可能是KFS中胶原沉积增多的机制之一。我们的结果表明miR-10a是一种很有前途的针对瘢痕疙瘩病变的新的有效策略。

We aimed to investigate expression profiles of microRNAs (miRNAs) in order to understanding the molecular mechanism(s) involved in the pathogenesis of keloid. We confirmed that miR-10a was significantly under-expressed in keloid fibroblasts (KFs) compared with normal dermal fibroblasts (NFs).Inhibition and induction of miR-10a respectively by miRNA inhibitor and retinoic acid (RA) in KFs and NFs revealed that IL-6 and pro-collagen type I (PICP) secretions in the culture supernatants were decreased by miR-10a inhibitor and increased by RA treatment. These results suggest that miR-10a regulates the synthesis and secretion of collagen in KFs and NFs. KF and NF cells were transfected with miR-10a oligonucleotide. The secretion of PICP was reduced in miR-10a transfected KFs and NFs. The miR-10a was significantly over-expressed in KFs and NFs after treatment with RA. The treated cells were harvested for miR-10a qPCR assay and the culture supernatant was analyzed for IL-6 and PICP secretion by ELISA method. The assays indicated that IL-6 concentrations were markedly increased in KFs compared with NFs after RA treatment.Our data confirmed the differential expression of miR-10a in KFs compared with NFs. The miR-10a regulates the synthesis and secretion of collagen in KFs and NFs. Down-regulation of miR-10a may be one of the mechanisms by which collagen is highly deposited in KFs. Our results showed that miR-10a is a promising new effective strategy for targeting keloid lesions.