miR-200c抑制人瘢痕疙瘩成纤维细胞增殖和胶原合成的研究

Inhibition studies of miR-200c on cell proliferation and collagen synthesis in human keloid fibroblasts

目的：探讨miR-200c对人瘢痕疙瘩成纤维细胞human keloid fibroblasts，HKFs）增殖和胶原合成的影响及阐明其机制。方法：将miR-200c mimics用oligofectami脂质体转染经转化生长因子（TGF）-β1诱导的HKFs，CCK-8法测细胞增殖变化；3H-脯氨酸掺入法测胶原蛋白水平的变化。Western blot检测细胞外信号调节激酶（extracellular signal-regulated kinase，ERK）及磷酸化ERK1／2蛋白表达变化。结果：在有或无TGF-β1刺激下，miR一200c均能明显抑制HKFs的增殖能力及胶原合成，并且能明显减低磷酸化ERK1／2蛋白表达水平，但对非磷酸化ERK表达无影响。结论：miR-200c能抑制HKFs增殖和胶原合成．并起拮抗TGF-B1的作用。其机制可能是通过降低ERK通路活性介导实现的。

Objectives： To investigate the effects of miR-200c on cell proliferation and collagen synthesis in human keloid fibmblasts （HKFs） and elucidate its mechanisms. Methods： HKFs were transfected with miR-200c mimics by oligofectami package. Cell proliferation was assessed by cell counting Kit-8; Rates of collagen synthesis were measured by 3H-proline incorporation assay. Changes of non-phosphorylated ERK and phosphorylated ERK1/2 protein expression were evaluated by Western blot. Results： miR-200e significantly inhibited cell proliferation and collagen synthesis in HKFs with or without TGF-β1 treatment. The expression of phosphorylated ERK1/2 was markedly reduced by miR-200c treatment but non-phos- phorylated ERK expression remained unchanged. Conclusion： miR-200c exhibits inhibitory effects on TGF-β1 and cell proliferation and collagen synthesis in HKFs, and its action mechanisms may be mediated by reducing the ERK pathway activity.