TGF-β1在人前列腺上皮细胞炎症过程中的表达机制及意义  

The expression mechanism and significance of TGF-β1 in the inflammatory process of human prostatic epithelial cells

作　　者：黄敏玉[1] 黄华武[1] 黄群[1] 吴军[1] 覃天资 孟冬冬[1] 龙毅[1] HUANG Minyu;HUANG Huawu;HUANG Qun;WU Jun;QIN Tianzi;MENG Dongdong;LONG Yi(Department of Urology,Affiliated Hospital of Youjiang Medical College for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院附属医院泌尿外科,广西百色533000

出　　处：《暨南大学学报（自然科学与医学版）》2020年第1期39-47,共9页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广西壮族自治区计划课题(Z20170222)。

摘　　要：目的:探讨炎症刺激诱导人前列腺上皮细胞中转化生长因子(TGF-β1)的表达水平,分析其影响机制及意义.方法:采用脂多糖诱导处理人前列腺上皮P69细胞构建炎症细胞模型,体外培养正常P69细胞(对照组)和炎症细胞(实验组)至对数生长期,运用酶联免疫法检测两组细胞中白细胞介素-6(IL-6)、IL-10和肿瘤坏死因子-α(TNF-α)的质量浓度.分别利用qPCR法和Western Blot法检测两组细胞中c-Myb和TGF-β1的mRNA表达和蛋白定量水平;应用Real time PCR测定两组细胞中hsa-miR-150的表达;采用hsa-miR-150-mimic和hsa-miR-150-inhibitor分别处理两组细胞,qPCR法和免疫荧光法观察c-Myb及TGF-β1的表达水平改变;采用c-Myb抑制剂处理后,观察两组细胞中TGF-β1的表达程度.结果:实验组IL-6、IL-10和TNF-α水平均较对照组明显升高(P<0.05),表明P69炎症细胞模型构建成功.实验组c-Myb和TGF-β1的蛋白表达水平较对照组增强;基线状态下实验组细胞hsa-miR-150、c-Myb和TGF-β1的mRNA表达水平均高于对照组(P<0.05);hsa-miR-150-mimic处理后,实验组hsa-miR-150、c-Myb和TGF-β1的mRNA表达水平与处理前比较无统计学差异(P>0.05),对照组hsa-miR-150、c-Myb和TGF-β1的mRNA表达水平较处理前水平升高(P<0.05),免疫荧光显示对照组c-Myb和TGF-β1的蛋白表达水平均高于实验组;hsa-miR-150-inhibitor处理后,实验组hsa-miR-150、c-Myb和TGF-β1水平较处理前明显降低(P<0.05),对照组hsa-miR-150、c-Myb和TGF-β1的mRNA表达水平与处理前比较无统计学差异(P>0.05),处理后两组间hsa-miR-150的mRNA表达结果比较无统计学差异(P>0.05),而两组间c-Myb和TGF-β1的mRNA表达水平分别比较均具有统计学差异(P<0.05),免疫荧光显示对照组c-Myb和TGF-β1的蛋白表达水平均低于实验组;c-Myb-inhibitor抑制处理前,两组细胞TGF-β1的mRNA表达水平存在显著性统计学差异(P<0.05);处理后,实验组TGF-β1的mRNA表达水平较处理前出现降低,对照组表Objective:To investigate the expression of transforming growth factor-β1(TGF-β1)in human prostatic epithelial cells induced by inflammatory stimulation,and to analyze its mechanism and significance.Methods:Inflammatory cell model was constructed by lipopolysaccharide-induced treatment of human prostatic epithelial P69 cells.Normal P69 cells(control group)and inflammatory cells(experimental group)were cultured in vitro until logarithmic growth stage.The level of interleukin-6,10(IL-6,10)and tumor necrosis factor-alpha(TNF-α)were detected by enzyme-linked immunosorbent assay.Protein and mRNA expression of c-Myb and TGF-β1 was detected by Western Blot and qPCR,respectively.Expression of hsa-microRNA-150 was detected by Real time PCR.After treated with hsa-microRNA-150 mimic and hsa-microRNA-150 inhibitor respectively,the expression level of c-Myb and TGF-β1 were observed by qPCR and immunofluorescence in two groups of cells.After treated with c-myb inhibitor,the expression of TGF-β1 in two groups was observed.Results:The level of IL-6,IL-10 and TNF-αin experimental group was significantly higher than those in control group(P<0.05),indicating successful establishment of the inflammatory cell model.The protein expression level of c-myb and TGF-β1 in experimental group was higher than that in control group.At baseline,the mRNA expression of hsa-mir-150,c-Myb and TGF-β1 in experimental group was higher in experimental group than that in control group(P<0.05).After hsa-mir-150-mimic treatment,there was no significant difference in the mRNA expression level of hsa-mir-150,c-Myb and TGF-β1 observed in experimental group when compared with that before treatment(P>0.05);while the mRNA expression level of hsa-miR-150,c-Myb and TGF-β1 in control group were higher than that before treatment(P<0.05).The protein expression level of c-Myb and TGF-β1 in control group was higher than that in experimental group when assayed by immunofluorescence method.After hsa-mir-150-inhibitor treatment,the level of hsa-mir-150,c-Myb a

关 键 词：转化生长因子-β1(TGF-β1) 脂多糖 前列腺炎症 表达机制 

分 类 号：R697[医药卫生—泌尿科学]