miR-384靶向HTRA1影响瘢痕疙瘩成纤维细胞的增殖和凋亡

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

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。

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.