瘢痕疙瘩成纤维细胞端粒酶活性及相关基因的表达

Telomerase activation and related gene expression of keloid fibroblasts

背景:研究发现人端粒酶催化亚单位在瘢痕疙瘩组织中有高表达。目的:拟进一步验证瘢痕疙瘩成纤维细胞中端粒酶活性及端粒酶催化亚单位、bcl-2mRNA在瘢痕疙瘩成纤维细胞中的表达及其在瘢痕疙瘩发生、发展中的意义。设计、时间及地点:单一样本观察,于2005-03/2006-01在解放军第二军医大学长海医院整形外科完成。对象:实验标本取自8例瘢痕疙瘩标本和8例正常皮肤标本。提供者对实验知情同意。方法:取瘢痕疙瘩边缘突起发红的部分,按组织块法培养原代成纤维细胞,取第三四代的细胞用于实验。主要观察指标:用PCR-ELISA方法测定瘢痕疙瘩和正常皮肤成纤维细胞端粒酶活性;用反转录-聚合酶链反应的方法检测培养的瘢痕疙瘩和正常皮肤成纤维细胞人端粒酶催化亚单位、bcl-2mRNA的表达情况。结果:①PCR-ELISA结果表明:瘢痕疙瘩成纤维细胞端粒酶活性明显高于正常皮肤成纤维细胞,其差异有非常显著性意义(P<0.01)。②反转录-聚合酶链反应结果显示:端粒酶催化亚单位和bcl-2mRNA在瘢痕疙瘩成纤维细胞中有较强的表达,在正常皮肤成纤维细胞中表达较弱,前者显著高于后者(P<0.01)。结论:瘢痕疙瘩成纤维细胞中端粒酶活性明显增高。推测人端粒酶催化亚单位mRNA在瘢痕疙瘩成纤维细胞中的高表达与端粒酶活性密切相关。

BACKGROUND： Previous study reveals that, telomerase catalytic subunit is highly expressed in keloid tissues. OBJECTIVE： To further verify the telomerase activation and the expression of human telomerase reverse transcriptase （hTERT） and bcl-2 mRNA in keloid fibroblast, and approach its role in the formation and development of keloid. DESIGN, TIME AND SETTING： A single sample observation was carded out in the Department of Plastic Surgery, Changhai Hospital, the Second Military Medical University of Chinese PLA （Shanghai, China） from March 2005 to January 2006. PARTICIPANTS： Eight keloid and normal skin were selected. All donators signed the informed consents. METHODS： Keloid tissues with prominences and redness were harvested. Subsequent to primary cultures, dermal fibroblasts at passages 3 to 4 were used in this study. MAIN OUTCOME MEASURES： The samples from 8 keloid fibroblast cell lines and 8 normal skin fibroblast cell lines were examined by using polymerase chain reaction-enzyme-linked immunoadsorbent assay （PCR-ELISA） for telomerase activation. The expression of hTERT and bcl-2 mRNA in the fibroblasts derived from keloid and normal skin tissue was detected by semi-quantitative reverse transcriptase-polymerase chain reaction （RT-PCR）. RESULTS： PCR-ELISA result showed the telomerase activation of keloid fibroblasts was extremely significantly higher than that of normal skin fibroblasts （P 〈 0.01）. RT-PCR analysis showed the expression of hTERT and bcl-2 mRNA was strong in keloid, whereas was weak in normal skin. The keloid fibroblasts revealed a significant higher expression compared to normal skin （P 〈 0.01 ）. CONCLUSION： The telomerase activation of keloid fibroblasts obviously hoists. High expression of hTERT mRNA in the fibroblasts of keloid may be related to the telomerase activation.