赛来昔布对瘢痕疙瘩成纤维细胞的影响及β_catenin、C_myc表达的作用2.doc

赛来昔布对瘢痕疙瘩成纤维细胞增殖的影响及β-catenin和C-myc表达的作用 宋凡伟 福建医科大学附属第二临床医院 ，朱世泽 通讯作者，Email:shizezhu@,福建医科大学附属第二临床医院，泉州医学高等专科学校 ，王朝阳，吴文艺，王韶华，李永峰，郑志芳 【摘要】目的：探讨赛来昔布对体外培养的瘢痕疙瘩成纤维细胞增殖的影响及β-catenin和C-myc表达的作用 方法 :用不同浓度（0umol/L、25umol/L、50umol/L、100umol/L、125 umol/L）及不同作用时间（24h、48h、72h）的赛来昔布作用于人瘢痕疙瘩成纤维细胞，CCK-8法检测人瘢痕疙瘩成纤维细胞的增殖活性，QRT-PCR、Western-blott检测体外培养的人正常皮肤及瘢痕疙瘩成纤维细胞中β-catenin和C-myc 的mRNA及蛋白表达。 结果: CCK-8法显示赛来昔布可显著抑制体外培养的人瘢痕疙瘩成纤维细胞增殖（P0.O5），呈剂量和时间依赖性；QRT-PCR和Western blot显示，体外培养的人瘢痕疙瘩成纤维细胞中，C-myc 及β-catenin蛋白的表达明显高于人正常皮肤成纤维细胞（P0.05），而β-catenin mRNA表达无显著差异,随着赛来昔布药物浓度的增加，在体外培养的人瘢痕疙瘩成纤维细胞中，β-catenin和C-myc蛋白及mRNA的表达逐渐降低，各组之间差别具有统计学意义（P0.05）。 结论：赛来昔布可能通过Wnt信号通路抑制体外培养的人瘢痕疙瘩成纤维细胞增殖，Wnt信号通路的异常是人瘢痕疙瘩发生的重要机制。 【关键词】 瘢痕疙瘩 赛来昔布 成纤维细胞 细胞增殖 The effect of Celecoxib on proliferation of Human Keloids Fibroblasts and the expression of β-catenin and in C-myc vitro 【Abstract】 Objective To investigate the effect of Celecoxib on proliferation of Human Keloids Fibroblasts and the expression of β-catenin and C-myc in vitro, to further explore the mechanism of it. Methods With different concentrations(0umol/L、25umol/L、50umol/L、100umol/L、125 umol/L) and effect time(24h、48h、72h) of Celecoxib on cultured keloid fibroblasts, and CCK-8 assay was adopted to evaluate cell survival，QRT-PCR and Western blot were performed to detect the expression ofβ-catenin and C-myc before and after intervention in cultured keloid fibroblasts, cultured normal fibroblasts. Result CCK-8 assay exibitted that Celecoxib could significantly inhibit the proliferation and activity of Human keloid fibroblasts with different concentrations(0umol/L、25umol/L、50umol/L、100umol/L、125 umol/L) and effect time(24h、48h、72h) (p0.05). RT-PCR and western blot results showed that the gene and protein expression of C-myc with the protein expression of β-catenin in cultured keloid fibroblasts were much higher than which in cultured normal fibroblasts（P0.05），with no the no difference of mRNA level ofβ-catenin （P0.05）；the mRNA and protein level of