炎症介质白细胞介素-8通过上调FTO抑制前列腺干细胞尿路上皮样分化  

Inflammatory Mediator Interleukin-8 Inhibits Urothelioid Differentiation of Prostate Stem Cells by Up-Regulating FTO

作　　者：孙江波 林婷婷[1,2,3] 李晓东[1,2,3] 许以城[1,2,3] 阮中天 魏勇 SUN Jiangbo;LIN Tingting;LI Xiaodong;XU Yicheng;RUAN Zhongtian;WEI Yong(Department of Urology,The First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China;Department of Urology,National Regional Medical Center,Binhai Campus of The First Affiliated Hospital,Fujian Medical University,Fuzhou 350212,China;Institute of Urology,Fujian Medical University,Fuzhou 350005,China)

机构地区：[1]福建医科大学附属第一医院泌尿外科,福州350005 [2]福建医科大学附属第一医院滨海院区国家区域医疗中心泌尿外科,福州350212 [3]福建医科大学泌尿外科研究所,福州350005

出　　处：《福建医科大学学报》2023年第6期435-440,共6页Journal of Fujian Medical University

基　　金：福建省卫生健康科技计划项目医学创新课题(2020CXA033);福建省财政专项项目(2020B018);福建省自然科学基金项目(2021J01232)。

摘　　要：目的 探讨炎症介质白细胞介素-8(IL-8)调控脂肪含量和肥胖相关基因(FTO)对前列腺干细胞(PSCs)尿路上皮样分化的影响。方法 从良性前列腺增生(BPH)患者手术标本组织中提取原代PSCs,与前列腺上皮细胞(RWPE-1)进行比较,通过特异性标志物CD49f进行鉴定。以Transwell共培养装置将PSCs与尿路上皮细胞共培养,诱导PSCs尿路上皮样分化。使用外源性IL-8处理PSCs。采用CCK-8实验检测PSCs细胞增殖能力;采用Transwell实验检测PSCs细胞迁移能力;采用Western-blot实验检测PSCs尿路上皮样分化的标志蛋白CK-18和UPK2,评价尿路上皮样分化程度,并检测FTO蛋白表达水平;采用免疫荧光检测敲低FTO后该标志蛋白表达水平变化。结果 CCK-8实验和Transwell实验结果均表明,IL-8可以抑制PSCs细胞的增殖和迁移能力。Western-blot实验结果显示,空白组、阳性对照组和IL-8处理组的CK-18相对表达量分别为(0.419±0.019)、(0.890±0.010)和(0.720±0.017),UPK2的相对表达量分别为(0.556±0.029)、(0.929±0.034)和(0.743±0.045),IL-8处理组的CK-18和UPK2的相对表达量较阳性对照组显著降低,差别有统计学意义(t=14.890,P<0.05;t=5.693,P<0.05)。空白组、阳性对照组和IL-8处理组的FTO蛋白相对表达量分别为(0.713±0.011)、(0.294±0.007)和(1.024±0.047),IL-8处理组均较阳性对照组显著升高,差别有统计学意义(t=26.630,P<0.05)。免疫荧光结果显示,IL-8+siRNA-FTO处理组和IL-8处理组的CK-18水平分别为(15.893±4.105)和(3.125±0.422),UPK2的水平分别(9.452±1.274)和(2.248±0.238),与单纯IL-8处理组比较,IL-8+siRNA-FTO处理组中CK-18的水平(t=16.044,P<0.05)和UPK2的水平(t=11.513,P<0.05)均显著升高,差别有统计学意义。结论 炎症介质IL-8通过上调FTO,抑制PSCs尿路上皮样分化。Objective This study aims to investigate the effects of fat mass and obesity-associated gene(FTO)regulated by interleukin-8(IL-8),on urothelioid differentiation of prostate stem cells(PSCs).Methods Primary PSCs were extracted from surgical specimens of patients with benign prostatic hyperplasia and identified using the specific marker CD49f,comparing them with human normal prostate epithelial cells(RWPE-1).PSCs were co-cultured with urothelial cells using Transwell co-culture chambers to induce urothelial-like differentiation in PSCs.Exogenous IL-8 was used to treat PSCs.The proliferation of PSCs was assessed using the CCK-8 assay,while the migration capability was evaluated through Transwell experiments.Western-blot analysis was employed to detect the expression of the urothelial-like differentiation markers CK-18 and UPK2 in PSCs and assess the degree of urothelial-like differentiation.Furthermore,Western-blot was used to determine FTO protein expression levels.Immunofluorescence was used to detect changes in the expression level of the markers after knocking down FTO.Results The results of both CCK-8 and Transwell experiments indicate that IL-8 can significantly inhibit the proliferation and migration abilities of PSCs.Western-blot experiment results show that the relative expression levels of CK-18 in the blank group,positive control group,and IL-8 treated group are(0.419±0.019),(0.890±0.010),and(0.720±0.017)respectively,and the relative expression levels of UPK2 are(0.556±0.029),(0.929±0.034),and(0.743±0.045)respectively.The CK-18 and UPK2 relative expression levels in the IL-8 treated group are significantly lower than those in the positive control group(t=14.890,P<0.05 and t=5.693,P<0.05).The relative expression levels of FTO in the blank group,positive control group,and IL-8 treated group are(0.713±0.011),(0.294±0.007),and(1.024±0.047)respectively.The FTO expression in the IL-8 treated group is significantly higher than that in the positive control group(t=26.630,P<0.05).Immunofluorescence result

关 键 词：白细胞介素-8 前列腺干细胞 尿路上皮样分化 脂肪含量和肥胖相关基因 

分 类 号：R697.3[医药卫生—泌尿科学]