人瘢痕疙瘩成纤维细胞中CASK／Idl通路的初步研究  

Preliminary study on CASK/Id1 pathway in fibroblasts of human keloid

作　　者：肖亮[1] 孙志波[1] 柯勇[1] 禹志宏[1] 贺光照[1] 任玉涵[1] 

机构地区：[1]湖北省十堰市人民医院创伤骨科,442000

出　　处：《中华整形外科杂志》2014年第2期105-111,共7页Chinese Journal of Plastic Surgery

摘　　要：目的验证人瘢痕疙瘩成纤维细胞中钙／钙调蛋白依耐性丝氨酸蛋白激酶（calcium／calmodulin dependent serine protein kinase，CASK）／分化抑制因子1（inhibitors of differentiation 1，Id1）通路的存在及意义。方法通过免疫荧光激光共聚焦证实瘢痕疙瘩和正常皮肤成纤维细胞中CASK和Idl蛋白表达和定位；RT—PCR和Western—blot分析瘢痕疙瘩和正常皮肤成纤维细胞中CASK和Idl的表达和差异；通过免疫沉淀验证瘢痕疙瘩成纤维细胞中CASK和Idl蛋白的天然结合。结果正常情况下体外培养的瘢痕疙瘩和正常皮肤成纤维细胞中均存在CASK和Idl蛋白的表达，CASK和Idl主要分布在成纤维细胞胞浆和胞核中；RT．PCR结果表明瘢痕疙瘩组CASKmRNA的表达量为0．658±0．024，低于正常对照组的1．0764±0．008（t=11．159，P〈0．05）；Id1的表达量为0．497±0．014，高于正常对照组的0．307±0．017（t=15．148，P〈0．05）；Western-blot结果表明瘢痕疙瘩组CASK蛋白的表达量为0．057±0．006，低于正常对照组的0．1684-0．012（t=13．524，P〈0．05）；Idl的表达量为0．8124±0．035，高于正常对照组的0．3684±0．031（t=16．356，P〈0．05）；免疫沉淀结果显示，CASK沉淀物中能检测到Id1，Id1沉淀物中能检测到CASK，CASK和Idl在瘢痕疙瘩成纤维细胞中存在天然结合。结论可能存在CASK／Id1信号通路参与了瘢痕疙瘩成纤维细胞的增殖，这与瘢痕疙瘩的发生存在一定联系。Objective To verify the existence and significance of calcium/calmodulin dependent serine protein kinase/inhibitors of differentiation 1 （ CASK/Id1 ） pathway in fibroblasts of human keloid. Methods Immunofluorescence laser was used to confirm CASK and Idl protein expression and localization in fibroblasts of the keloid and normal skin. RT-PCR and Western-blot were adopted to analysis the CASK and Idl expression and differences between keloid and normal skin fibroblasts. The natural combination of CASK and Idl protein of keloid fibroblasts was tested by immunoprecipitation. Results CASK and Idl protein expression were both found in fibroblast cells of keloid and normal skin under normal circumstances. Most of CASK and Idl were distributed in the cytoplasm and nucleus of fibroblasts. The results of RT-PCR showed that the expression of CASK mRNA in the keloid group was 0. 658 ± 0. 024, which was lower than that in the normal control group（1.076 ± 0.008, t =11.159, P〈0.05）. The expression of Idl mRNA was 0. 497 ± 0. 014, which was higher than that in the normal control group （0. 307 ±0. 017, t = 15. 148,P 〈0. 05）. The results of Western-blot showed that the expression level for CASK protein in the keloid group was 0. 057± 0. 006, which was lower than that in the normal control group （0. 168 ± 0. 012, t = 13. 524,P 〈 0. 05 ） ; the expression of Idl protein was 0. 812 ± 0. 035, which was higher than that in the normal control group （0. 368 ± 0. 031, t = 16. 356, P 〈 0. 05 ）. The results of immunoprecipitation showed that Id1 could be detected in the CASK precipitate, while CASK also could be detected in the Idl precipitate. There was a natural binding of CASK and Idl in keloid fibroblasts. Conclusion CASK/Id1 signal pathway may be existed and involved in the proliferation of keloid fibroblasts, which is related with the occurrence of keloid.

关 键 词：瘢痕疙瘩 成纤维细胞 分化抑制因子1 免疫沉淀法 

分 类 号：R622[医药卫生—整形外科]