| 章杰 李文芳 王芳 杨红华 万芸 刘伟.长链非编码RNA H19对瘢痕疙瘩成纤维细胞增殖的影响[J].实用中西医结合临床,2013,(9):1-3,9 |
| 长链非编码RNA H19对瘢痕疙瘩成纤维细胞增殖的影响 |
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| DOI： |
| 中文关键词: 瘢痕疙瘩 长链非编码RNA H19 增殖 |
| 英文关键词: Keloid IncRNA H19 Proliferation |
| 基金项目:江西省教育厅科学技术研究项目资助（编号GJJ13153） |
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| 中文摘要: |
| 目的：探讨长链非编码RNA H19在瘢痕疙瘩组织中的表达情况及对瘢痕疙瘩成纤维细胞增殖的影响及机制。方法：本次实验共收集8例瘢痕疙瘩患者组织标本，对照组取自8例成熟皮肤、8例成熟瘢痕，通过RT-PCR法检测长链非编码RNA H19的表达；对体外培养的人瘢痕疙瘩成纤维细胞，首先采用脂质体介导的scramble siRNA、H19 siRNA、阴性对照siRNA转染体外培养的瘢痕疙瘩成纤维细胞，验证H19 siRNA的转染效率；MTT法检测转染H19 siRNA后成纤维细胞的增殖，Westing blotting检测转染后血管内皮生长因子(vascular endothelial growth factor,VEGF）和雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)的表达。结果：与对照组相比，H19 mRNA的表达量在瘢痕疙瘩患者中明显升高（P <0.05）；与scramble siRNA、阴性对照siRNA相比，转染48 h后H19 siRNA转染的成纤维细胞H19 mRNA表达量明显降低（P <0.05）；与空白对照组（转染时不加siRNA和脂质体）和阴性对照组siRNA组相比，使用H19的siRNA转染后，成纤维细胞的增殖明显受到抑制，mTOR和VEGF的表达水平明显降低（P <0.05）。结论：下调非编码长链RNA H19的表达量可以抑制瘢痕疙瘩成纤维细胞的增殖。 |
| 英文摘要: |
| Objective: To investigate the effect of long non-coding RNA H19 on proliferation of human keloid fibroblasts and the possible role of the mechanisms underlying keratinocyte proliferation. Methods: The samples were collected from 24 patients, in which were 8 with keliods, 8 with normal scars and 8 with normal skin control. The H19 expression level in human keloid fibroblasts, normal scars and normal skin were determined by RT-PCR. H19-siRNA, scramble siRNA and control siRNA mediated by lipfectamin were introduced into human keloid fibroblasts, respectively. The silencing effect of H19 was verified by RT-PCR. An effect of H19 on cell proliferation was investigated by MTT colorimetric assay. To evaluate which signaling mediators were regulated by H19 in keloid fibroblast, the expression of mammalian target of rapamycin(mTOR) and vascular endothelial growth factor (VEGF) were tested by western blot. Results:The data showed that H19 levels were markedly increased in human keloids ?broblasts compared with normal controls（P <0.05）. Forty-eight hours after transfection, mRNA expression of H19 in H19 siRNA treated cells was remarkedly reduced as compared with those in the scramble siRNA and control siRNA treated cells(P <0.05).H19 siRNA treatment obviously inhibit the proliferation of Keloid Fibroblasts (P <0.05).The study further veri?ed that H19 was associated with mTOR and VEGF, and that this association resulted in the proliferation of human keloids ?broblasts. Conclusion: These data suggest that down-regulating the expression of H19 strongly suppressed cell proliferation of keloid fibroblasts. |
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