circ_0015756靶向miR-136调控银屑病角质形成细胞增殖和凋亡的机制研究

circ_0015756 Regulates the Mechanism of Psoriasis Keratinocyte Proliferation and Apoptosis by Targeting miR-136

目的探讨环状_0015756(circ_0015756)对银屑病角质形成细胞增殖和凋亡的影响及其分子机制。方法采用实时荧光定量聚合酶链反应(qRT-PCR)法检测正常皮肤组织、银屑病患者皮损组织中circ_0015756与微小核糖核酸-136(miR-136)的表达量;原代分离培养人银屑病角质形成细胞,根据转染物不同将细胞分为以下几组:circ_0015756小分子干扰RNA(si-circ_0015756)组、si-circ_0015756阴性对照(si-NC)组、miR-136寡核苷酸模拟物(miR-136)组、miR-136寡核苷酸模拟物的阴性对照(miR-NC)组、si-circ_0015756+miR-136特异性寡核苷酸模拟物的阴性对照(anti-miR-NC)组、si-circ_0015756+miR-136特异性寡核苷酸抑制剂(anti-miR-136)组;细胞计数试剂盒-8(CCK-8)、平板克隆形成实验与流式细胞术分别检测细胞增殖、克隆形成能力及凋亡率;双荧光素酶报告基因实验检测circ_0015756与miR-136的靶向关系。结果与正常皮肤组织比较,银屑病患者皮损组织中circ_0015756的相对表达量升高(P<0.05),miR-136的相对表达量降低(P<0.05);与si-NC组比较,si-circ_0015756组细胞活力降低(P<0.05),克隆形成数减少(P<0.05),凋亡率升高(P<0.05);circ_0015756可靶向调控miR-136;与miR-NC组比较,miR-136组细胞活力降低(P<0.05),克隆形成数减少(P<0.05),凋亡率升高(P<0.05);与si-circ_0015756+anti-miRNC组比较,si-circ_0015756+anti-miR-136组细胞活力升高(P<0.05),克隆形成数增多(P<0.05),凋亡率降低(P<0.05)。结论干扰circ_0015756表达可通过靶向调控miR-136而抑制银屑病角质形成细胞增殖、克隆形成及诱导细胞凋亡。

Objective In order to explore the effects of circ_0015756 on the proliferation and apoptosis of psoriasis keratinocytes and its molecular mechanism.Methods Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR)method was used to detect the expression of circ_0015756 and miR-136 in normal skin tissues and skin lesions of patients with psoriasis.The primary isolation and culture of human psoriatic keratinocytes were divided into the following groups according to the different transfectants:circ_0015756 small interfering RNA(si-circ_0015756)group,si-circ_0015756 negative control(si-NC)group,miR-136 oligonucleotide mimic(miR-136)group,miR-136 oligonucleotide mimic negative control(miR-NC)group,si-circ_0015756+anti-miR-NC group and si-circ_0015756+anti-miR-136 group.Cell counting kit-8(CCK-8),plate clone formation experiment and flow cytometry were used to detect cell proliferation,clone formation ability and apoptosis rate.The dual luciferase reporter gene experiment was used to detect the relationship between circ_0015756 and miR-136.Results Compared with normal skin tissues,the expression of circ_0015756 in the skin lesions of patients with psoriasis was increased(P<0.05),while the expression of miR-136 was decreased(P<0.05).Compared with the si-NC group,the si-circ_0015756 group showed reduced cell viability(P<0.05),the number of clones was reduced(P<0.05),while the apoptosis rate was increased(P<0.05).circ_0015756 could target miR-136.Compared with the miR-NC group,the cell viability of the miR-136 group was decreased(P<0.05),the number of clone formation was decreased(P<0.05),while the apoptosis rat was increased(P<0.05).Compared with the si-circ_0015756+anti-miR-NC group,the cell viability of the sicirc_0015756+anti-miR-136 group was increased(P<0.05),the number of clones was increased(P<0.05),while the apoptosis rate was decreased(P<0.05).Conclusion Interference with the expression of circ_0015756 could inhibit the proliferation,clone formation and induce apoptosis of psoriatic keratinocytes by targeted regulation of miR-136.