荧光定量分析人皮肤瘢痕疙瘩组织中miRNA-34s的表达

Fluorescent quantitative analysis on the expression of miRNA-34s in human skin keloid tissue

背景:通过分析不同肿瘤组织与正常组织miRNA表达谱的差异,有可能筛选到特异性好、灵敏度高、可作为特定肿瘤分子标志物的miRNA。目的:采用实时荧光定量RT-PCR检测miRNA-34家族(miRNA-34s:miR-34a/b/c)在瘢痕疙瘩和正常皮肤组织中的差异表达,分析评价microRNA-34s在瘢痕疙瘩形成发展的作用及可能的机制影响。方法:收集手术切除瘢痕疙瘩组织(10例)和正常皮肤(2例);TRIZOL法提取标本的总RNAs并进行质检,再采用Ambion′s miRNA Isolation Kit从总RNAs中分离microRNA,采用实时荧光定量RT-PCR技术检测miRNA-34s(miR-34a/b/c)在瘢痕疙瘩组织和正常皮肤组织中的表达水平。结果与结论:miRNA-34s(miRNA-34a/b/c)在瘢痕疙瘩组织中均呈下调表达(P<0.01)。表明该家族成员参与了瘢痕疙瘩的形成发展,下调表达的miRNA-34s可能导致了瘢痕疙瘩的肿瘤性生长。

BACKGROUND：Understanding the difference of miRNA-34s expression in normal tissue and tumor tissue wil contribute to screen out a miRNA with high sensitivity as the specific tumor molecular marker.OBJECTIVE：To investigate the differential expression of miRNA-34s （miR-34a/b/c） between normal skin and keloid tissue using real-time fluorescent quantitative PCR, and to evaluate the role and mechanisms of miRNA-34s in keloid formation and development.METHODS：Ten cases of keloid tissue and two cases of normal skin tissue were col ected as specimens. Total RNAs were extracted from keloid and nomal skin tissue by Trizol method, and miRNA-34s were further isolated by Ambion’s miRNA Isolation Kit. Real-time fluorescent quantitative PCR was applied to verify expression levels of microRNA-34s （miR-34a/b/c） in keloid tissue and normal skin tissue.RESULTS AND CONCLUSION：miRNA-34s （miRNA-34a/b/c） expression was down-regulated in keloid tissue compared with normal skin tissue （P〈0.01）. The findings showed that miRNA-34s （miRNA-34a/b/c） are involved in keloid formation and development, and down-regulation of the family member may result in neoplastic growth of keloid.