实用医学杂志 ›› 2021, Vol. 37 ›› Issue (2): 226-232.doi: 10.3969/j.issn.1006⁃5725.2021.02.019

• 临床研究 • 上一篇    下一篇

长链非编码lncRNA DBH⁃AS1对瘢痕疙瘩成纤维细胞凋亡的影响及作用机制

贾春英, 张荣明
  

  1. 锦州医科大学附属第三医院烧伤整形外科(辽宁锦州121000)
  • 出版日期:2021-01-25 发布日期:2021-01-25
  • 通讯作者: 张荣明 E⁃mail:zrm99999@126.com
  • 基金资助:
    辽宁省自然科学基金资助项目(编号:2014022035)辽宁省科技厅联合基金项目(编号:2015020351)

Impact of long non⁃coding lncRNA DBH⁃AS1 on apoptosis of keloid fibroblasts and its mechanism of action

JIA Chunying,ZHANG Rongming
  

  1. Department of Burn and Plastic Surgery,Affiliated Third Hospital,Jin⁃zhou Medical University,Jinzhou 121000,China
  • Online:2021-01-25 Published:2021-01-25
  • Contact: ZHANG Rongming E⁃mail:zrm99999@126.com

摘要:

目的 探讨长链非编码 lncRNA DBH⁃AS1(DBH⁃AS1)对瘢痕疙瘩成纤维细胞凋亡的影响及 机制。方法 采用实时荧光定量逆转录聚合酶链反应(qRT⁃PCR)检测瘢痕疙瘩和正常皮肤组织中 DBH⁃ AS1、MicroRNA⁃138(miR⁃138)和 Hypoxia⁃inducible factor⁃1α(HIF⁃1α)的 mRNA 表达水平。采用双荧光素酶 报告分析三者的结合情况。采用流式细胞技术、qRT⁃PCR 和免疫印迹法(Western blot)检测敲低或过表达 DBH⁃AS1 对瘢痕疙瘩成纤维细胞凋亡的影响,并分析 miR⁃138 HIF⁃1α的表达变化。结果 DBH⁃AS1 HIF⁃1α瘢痕疙瘩组织和细胞中显著上调,而 miR⁃138 明显下调(P < 0.05)。双荧光素酶结果显示 DBH⁃AS1 能够靶向结合 miR⁃138,miR⁃138 能够靶向结合 HIF⁃1α。低表达 DBH⁃AS1 或过表达 miR⁃138 明显促进瘢痕 疙瘩成纤维细胞凋亡。敲低 miR⁃138 能逆转沉默 DBH⁃AS1 对瘢痕疙瘩成纤维细胞凋亡的影响,敲低 HIF⁃ 1α显著抑制沉默 miR⁃138 对细胞凋亡的作用(P < 0.05)。结论 低表达 DBH⁃AS1 能够通过调控 miR⁃138/ HIF⁃1α通路促进瘢痕疙瘩成纤维细胞凋亡,提示DBH⁃AS1

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Abstract:

Objective To explore the effect of long non⁃coding lncRNA DBH⁃AS1(DBH⁃AS1)on apoptosis of keloid fibroblasts and its mechanism of action. Methods Real⁃time fluorescent quantitative reverse transcription polymerase chain reaction(qRT⁃PCR)was used to detect mRNA expression levels of DBH⁃AS1,microRNA⁃138 (miR⁃138)and Hypoxia⁃inducible factor⁃1α(HIF⁃1α)in keloids and normal skin tissues. Dual Luciferase Report was applied to analyze the combination of the three proteins. Flow cytometry,qRT ⁃PCR and Western blot were utilized to detect the effect of knockdown or overexpression of DBH⁃AS1 on apoptosis of keloid fibroblasts,the expression changes of miR⁃138 and HIF⁃1α were also analyzed. Results DBH⁃AS1 and HIF⁃1α were significantly up⁃regulated in keloid tissues and cells while miR⁃138 was significantly down⁃regulated(P < 0.05). The Dual Lucif⁃ erase Report verified that DBH⁃AS1 could have targeted binding with miR⁃138,and miR⁃138 could have targeted binding with HIF⁃1α in keloid fibroblast. Low expression of DBH⁃AS1 or overexpression of miR⁃138 significantly promoted apoptosis of keloid fibroblasts. Knockdown of miR⁃138 could reverse the effect of silencing DBH⁃AS1 on apoptosis of keloid fibroblasts,and knockdown of HIF⁃1α significantly inhibited the effect of silencing miR⁃138 on apoptosis(P < 0.05). Conclusions Silencing DBH⁃AS1 promoted apoptosis of keloid fibroblasts by regulating the miR⁃138/HIF⁃1α pathway,

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