抑制miR-31表达对人角质形成细胞增殖、凋亡的影响研究  

Effect of inhibiting the miR-31 expression on the proliferation and apoptosis of human keratinocyte cells

作　　者：石娴[1] 石年[1] SHI Xian;SHI Nian(Edong Medical Group Huangshi Central Hospital,Huangshi Hubei 435000,China)

机构地区：[1]鄂东医疗集团黄石市中心医院,湖北黄石435000

出　　处：《蚌埠医学院学报》2020年第3期315-319,共5页Journal of Bengbu Medical College

摘　　要：目的:探讨miR-31对人角质形成细胞系HaCaT细胞增殖和凋亡的影响以及可能机制。方法:培养HaCaT细胞,利用瞬时转染的方法,对不同组细胞进行转染。反义寡核苷酸技术(ASO)组:转染微小RNA(miR)-31 ASO;对照ASO组:转染对照ASO;空白对照组:转染空白质粒。MTT法检测不同转染组细胞增殖情况,Annexin V-FITC/PI双染色检测不同转染组细胞凋亡情况,PI单染色法检测不同转染组细胞周期,Western blotting检测不同转染组细胞中Rho相关结构域BTB蛋白质1(RhoBTB1)蛋白表达。结果:ASO组细胞中miR-31表达量明显低于对照ASO组和空白对照组(P<0.01);ASO组细胞在24、48、72、96 h时吸光度值均低于对照ASO组和空白对照组(P<0.05~P<0.01),且3组细胞随着时间推移,吸光度值均较之前逐渐增加(P<0.01);ASO组细胞凋亡率、G0/G1期细胞比例、RhoBTB1蛋白表达量明显高于对照ASO组和空白对照组(P<0.01);而S期和G2期细胞比例明显低于对照ASO组和空白对照组(P<0.01)。结论:miR-31可能参与了人角质形成细胞系HaCaT细胞增殖、凋亡过程,可能通过调控RhoBTB1蛋白表达而实现这一生物学过程。Objective:To investigate the effects of inhibiting the miR-31 expression on the proliferation and apoptosis of human keratinocyte cells,and its mechanism.Methods:The HaCaT cells were cultured,and transfected using transient transfection method.These cells were divided into the antisense oligonueleotide(ASO)group(treatment with miR-31 ASO),control ASO group(treatment with control ASO)and blank control group(treatment with blank plasmid).The cell proliferation,apoptosis,cell cycle and Rho related domain BTB protein 1(RhoBTB1)expression in three groups were detected using MTT assay,Annexin V-FITC/PI double staining,PI single staining and Western blotting,respectively.Results:The expression level of miR-31 in ASO group were significantly lower than that in ASO control group and blank control group(P<0.01).The absorbance values in ASO group at 24 h,48 h,72 h and 96 h were significantry lower than that in ASO control group and blank control group(P<0.05 to P<0.01),and the absorbance values in three groups gradually increased as time went on(P<0.01).The apoptotic rate,proportion of G0/G1 phase cells and expression level of RhoBTB1 protein in ASO group were significantly higher than those in ASO control group and blank control group(P<0.01),while the proportion of S/G2 phase cells in ASO group were significantly lower than those in ASO control group and blank control group(P<0.01).Conclusions:miR-31 may be involved in the proliferation and apoptosis of human keratinocyte cell lines HaCaT cells,which may achieve this biological process by regulating the expression of RhoBTB1 protein.

关 键 词：微小RNA 人角质形成细胞 反义寡核苷酸技术 细胞增殖 细胞凋亡 

分 类 号：R752.53[医药卫生—皮肤病学与性病学]