Zhang Jie, Liu Cai Yue, Wan Yun, Peng Li, Li Wen Fang, Qiu Jia Xuan
Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Department of Plastic Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China.
Oncol Lett. 2016 Oct;12(4):2835-2839. doi: 10.3892/ol.2016.4931. Epub 2016 Aug 2.
The expression of the long non-coding RNA (lncRNA) H19 is associated with proliferation in tumors. In order to investigate whether H19 may additionally mediate the proliferation of fibroblasts in human keloid disease, the present study collected samples from 24 subjects, including 8 with keloids, 8 with normal scars and 8 normal skin controls. Reverse transcription-polymerase chain reaction revealed that H19 levels were markedly increased in human keloids compared with normal scars and normal skin controls (P=0.017). In order to identify a potential role for H19 in the proliferative activity of human keloid fibroblasts, small interfering (si)RNA-mediated silencing experiments were performed. H19 siRNA treatment markedly inhibited the proliferation of keloid fibroblasts, as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (P=0.008). In order to identify the signaling mediators that are regulated by H19 in keloid fibroblasts, the expression levels of mammalian target of rapamycin (mTOR) and vascular endothelial growth factor (VEGF) were examined using western blotting. The results confirmed that knockdown of H19 inhibited mTOR and VEGF expression. In summary, the results indicate that H19 may be associated with increased proliferative activity of keloid fibroblasts, which may be mediated by mTOR and VEGF.
长链非编码RNA(lncRNA)H19的表达与肿瘤增殖相关。为了研究H19是否还可能介导人类瘢痕疙瘩疾病中成纤维细胞的增殖,本研究收集了24名受试者的样本,其中包括8名瘢痕疙瘩患者、8名正常瘢痕患者和8名正常皮肤对照者。逆转录-聚合酶链反应显示,与正常瘢痕和正常皮肤对照相比,人类瘢痕疙瘩中H19水平显著升高(P = 0.017)。为了确定H19在人类瘢痕疙瘩成纤维细胞增殖活性中的潜在作用,进行了小干扰(si)RNA介导的沉默实验。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐检测评估,H19 siRNA处理显著抑制了瘢痕疙瘩成纤维细胞的增殖(P = 0.008)。为了确定在瘢痕疙瘩成纤维细胞中受H19调控的信号介质,使用蛋白质印迹法检测了雷帕霉素靶蛋白(mTOR)和血管内皮生长因子(VEGF)的表达水平。结果证实,H19的敲低抑制了mTOR和VEGF的表达。总之,结果表明H19可能与瘢痕疙瘩成纤维细胞增殖活性增加有关,这可能由mTOR和VEGF介导。