中国麻风皮肤病杂志 ›› 2021, Vol. 37 ›› Issue (4): 198-203.doi: 10.12144/zgmfskin202104198

• 论著 • 上一篇    下一篇

miR-384靶向HTRA1影响瘢痕疙瘩成纤维细胞的增殖和凋亡

王东,王珍,刘岩   

  1. 沈阳市第七人民医院皮肤科，辽宁沈阳，110003
  • 出版日期:2021-04-15 发布日期:2021-04-07
  • 通讯作者: 王东，E-mail: DRwangdong123@126.com

miR-384 targeting HTRA1 affects the proliferation and apoptosis of keloid fibroblasts

WANG Dong, WANG Zhen, LIU Yan   

  1. Dermatology Department, The Seventh People's Hospital of Shenyang, Shenyang 110003, China
  • Online:2021-04-15 Published:2021-04-07
  • Contact: WANG Dong, E-mail: DRwangdong123@126.com

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摘要： 目的：明确miR-384对瘢痕疙瘩成纤维细胞(KFBs)增殖和凋亡的影响。方法：RT-qPCR和Western Blot检测miR-384和靶向高温需求因子A1（HTRA1）在KFBs和人瘢痕疙瘩皮肤组织中的表达。CCK-8细胞计数试剂盒检测KFBs增殖活力；流式细胞术检测KFBs细胞凋亡；Western Blot检测Ki-67、增殖细胞核抗原（PCNA）、B淋巴细胞瘤-2（Bcl-2）和Bcl-2相关X蛋白（Bax）表达。双荧光素酶报告基因实验验证miR-384对HTRA1的靶向作用。结果：KFBs和人瘢痕疙瘩皮肤组织中miR-384呈低表达，HTRA1呈高表达。转染anti-miR-384后KFBs增殖活力升高，细胞凋亡率降低，Ki-67、PCNA和Bcl-2蛋白表达升高，Bax蛋白表达降低。转染si-HTRA1后KFBs增殖活力降低，细胞凋亡率增加，Ki-67、PCNA和Bcl-2蛋白表达降低，Bax蛋白表达升高。miR-384靶向负性调控HTRA1表达。结论：miR-384可能通过靶向调控HTRA1促进瘢痕疙瘩成纤维细胞增殖，抑制细胞凋亡。

关键词: miR-384, HTRA1, 瘢痕疙瘩, 细胞增殖, 凋亡

Abstract: Objective: To determine the effect of miR-384 on the proliferation and apoptosis of keloid fibroblasts (KFBs). Methods: The levels of miR-384 and targeting high temperature requirement A1 (HTRA1) in KFBs and human keloid skin tissue were detected by RT-qPCR and Western blot. The proliferation activity of KFBs was detected by CCK-8. The KFBs apoptosis was detected by flow cytometry. The expression levels of Ki-67, proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) were detected by Western Blot. The targeting effect of miR-384 on HTRA1 was detected by double luciferase reporter gene experiment. Results: The expressions of miR-384 in KFBs and human keloid skin tissues showed low levels, while that of HTRA1 showed high level. After transfection with anti-miR-384, the proliferation activity of KFBs was increased, the apoptosis rate was decreased, the expression level of Ki-67, PCNA and Bcl-2 protein was increased, and that of Bax protein was decreased. After transfection with si-HTRA1, the proliferation activity of KFBs was decreased, the apoptosis rate was increased, the expression of Ki-67, PCNA and Bcl-2 protein was decreased, and the expression of Bax protein was increased. The targeted-regulating of miR-384 on HTRA1 expression was negative. Conclusion: miR-384 can promote proliferation and inhibit apoptosis of the KFBs through targeted regulating of HTRA1.

Key words: miR-384, HTRA1, keloid, cell proliferation, apoptosis