miRNA-199a-5p在热损伤人皮肤成纤维细胞中的表达及其功能研究

Expression and function of miRNA-199a-5p in human skin fibroblasts after thermal injury

目的:观察热损伤人皮肤成纤维细胞中miRNA-199a-5p表达量的变化,以及对皮肤成纤维细胞增殖、迁移、侵袭功能的影响。方法:分别用人皮肤成纤维细胞株(HSF)隔水52℃水浴30 s、60 s制造细胞热损伤模型,对比观察细胞热损伤状态;用四甲基偶氮唑盐(MTT)检测热损伤24 h、48 h后细胞的增殖率;用qRT-PCR检测热损伤24 h、48 h后细胞miRNA-199a-5p表达量的变化情况;用划痕和Transwell测试细胞的迁移、侵袭功能;用模拟物提高热损伤细胞中miRNA-199a-5p的表达量,检测热损伤细胞的增殖和侵袭情况。结果:52℃30 s可以很好地制造人皮肤成纤维热损伤模型;与对照组比较,热损伤后miRNA-199a-5p表达量下调[24 h,(2.67±0.35)%,P<0.001;48 h,(18.42±0.44)%,P<0.001],细胞增殖显著受到抑制[24 h,(17.10±3.21)%,P<0.002;48 h,(25.93±1.74)%,P<0.001],细胞的迁移功能显著下降[(35.35±7.07)%,P<0.001],侵袭功能亦被显著抑制[(57.14±9.52)%,P<0.007]。通过模拟物提高热损伤细胞中miRNA-199a-5p的表达量后,细胞的增殖显著上调[(136.55±6.14)%,P<0.003],细胞的侵袭能力增强[(196.88±15.63)%,P<0.003]。结论:热损伤人皮肤成纤维细胞中miRNA-199a-5p的表达量与细胞增殖、迁移和侵袭正相关,提高热损伤细胞中miRNA-199a-5p的表达量可以帮助热损伤细胞快速转归。

Objective:This study aims to observe the changes of miRNA-199 a-5 p expression in human skin fibroblasts with thermal injury and its effects on the proliferation,migration and invasion of skin fibroblasts. Methods:Cell models of thermal injury were established using human skin fibroblast cell line by bathing in water at 52℃ for 30 s and 60 s,respectively,and the thermal injury of cells was observed and compared. The proliferation rate of cells 24 h and 48 h after thermal injury was detected by MTT assay. The changes of miRNA-199 a-5 p expression in the cells 24 and 48 h after thermal injury were determined through q RT-PCR. Cell migration and invasion were detected using wound-healing assay and Transwell assay. The expression of miRNA-199 a-5 p in thermally injured cells was increased using mimics,and the proliferation and invasion of thermally injured cells were detected. Results:Bathing in water at 52 ℃ for 30 s could successfully establish a thermal injury model in human skin fibroblasts. After thermal injury,miRNA-199 a-5 p expression was down-regulated[24 h,(2.67±0.35)%,P<0.001;48 h,(18.42±0.44)%,P<0.001],cell proliferation was significantly inhibited[24 h,(17.10±3.21)%,P<0.002;48 h,(25.93±1.74)%,P<0.001],cell migration decreased significantly[(35.35±7.07)%,P<0.001],and cell invasion was also significantly inhibited[(57.14±9.52)%,P<0.007]. After enhancing miRNA-199 a-5 p expression in the thermally injured cells using mimics,the proliferation of the cells increased significantly[(136.55±6.14)%,P<0.003],and the invasion ability of the cells was enhanced[(136.55±6.14)%,P<0.003]. Conclusion:The expression level of miRNA-199 a-5 p in human skin fibroblasts with thermal injury is positively correlated with cell proliferation,migration and invasion.Increasing the expression level of miRNA-199 a-5 p in thermally injured cells can help it rapid return.