下调miR-21通过PTEN/PI3K/Akt信号转导抑制人增生性瘢痕成纤维细胞增殖的实验研究

Down-regulation of miR-21 inhibits the proliferation of human hypertrophic scar fibroblast cells via PTEN/PI3K/Akt signal transduction

目的探讨miR-21与PTEN相互作用关系及对PI3K/Akt信号通路和增生性瘢痕成纤维细胞增殖的影响,以期为临床诊断治疗增生性瘢痕提供新靶点。方法分别用qPCR和Western blot检测增生性瘢痕组织和细胞中miR-21和PTEN表达;双荧光素酶报告基因检测实验验证miR-21与PTEN调控关系;Western blot检测miR-21对HSF细胞中PTEN蛋白表达的影响;MTT检测细胞增殖能力的变化。结果在增生性瘢痕组织和细胞中miR-21普遍高表达,PTEN普遍低表达;双荧光素酶报告实验和Western blot证实了miR-21对PTEN的靶向调控作用;下调miR-21可通过上调PTEN表达来抑制PI3K/Akt通路活性进而抑制HSF细胞增殖。结论下调miR-21可通过PTEN/PI3K/Akt信号转导抑制人增生性瘢痕成纤维细胞增殖,显示出其诊断治疗增生性瘢痕的潜能,并可能为临床治疗提供新靶点。

Objective To investigate the relationship between miR-21 and PTEN as well as the effect of miR-21 on the PI3K/Akt signaling pathway and proliferation of hypertrophic scar fibroblasts cells in order to provide a new target for clinical diagnosis and treatment of hypertrophic scars.Methods The expressions of miR-21 and PTEN in hypertrophic scar tissues and cells were detected by qPCR and Western blot,respectively.The dual luciferase reporter assay was used to detect the relationship between miR-21 and PTEN.Western blot was used to detect the expression of PTEN protein.The influence of miR-21 on cell proliferation was detected by MTT assay.Results miR-21 was generally up-expressed in hypertrophic scar tissues and cells,while PTEN generally had low expression.Dual luciferase reporter assay and Western blot confirmed the regulation of miR-21on PTEN;down-regulation of miR-21 could up-regulate PTEN level,which then inhibited PI3K/Akt pathway and HSF cell proliferation.Conclusion Down-regulating miR-21 can inhibit the proliferation of human hypertrophic scar fibroblasts through PTEN/PI3K/Akt signal pathway,which shows its potential capacity for diagnosis and treatment of hypertrophic scars and may provide a new target for clinical therapy.