THP-1细胞对体外培养的人视网膜色素上皮细胞c-fos表达的上调作用  

Up-regulation of c-fos in cultured human retinal pigment epithelial cells by co-culturing with THP-1 cells

作　　者：王静波[1] 惠延年[2] 关娟[1] 杨丽[3] 

机构地区：[1]解放军总医院第二附属医院眼科,北京100091 [2]第四军医大学西京医院眼科,陕西西安710032 [3]山东省济南师范学校,山东济南250031

出　　处：《眼视光学杂志》2009年第2期110-113,共4页Chinese Journal of Optometry & Ophthalmology

基　　金：西安市科技攻关项目(GG05163)

摘　　要：目的检测与单核/巨噬细胞株THP-1细胞共培养的人视网膜色素上皮(retinal pigment epithelium,RPE)细胞c-fos的表达。方法在Transwell小室中共培养THP-1细胞和人RPE细胞达不同的时间(0,2h,3h,24h,48h,72h),采用免疫荧光染色法和逆转录聚合酶链反应(RT-PCR)观察RPE细胞c-fos蛋白和mRNA的表达变化。结果在未与THP-1细胞共培养的RPE细胞中,c-fos表达微弱或不表达,细胞浆和部分细胞核为浅黄绿色荧光。与THP-1细胞共培养达2h时,RPE细胞的荧光强度逐渐增强并出现核移位。随着与THP-1细胞共培养时间的延长,RPE细胞c-fosmRNA的表达(0h,0.40±0.07)开始逐渐增强,于2h时达峰值(2h,0.91±0.10,P=0.002),随后表达减弱,至3h时(3h,0.51±0.08)降至近未共培养时的水平。结论与THP-1细胞共培养可在短期内迅速上调体外培养的人RPE细胞c-fos蛋白和mRNA的表达,且蛋白表达出现核移位现象,提示THP-1细胞可引起RPE细胞c-fos的活化。Objective To study the effects of co-culturing with THP-1 on the expression of c-fos in human retinal pigment epithelium （RPE） cells in vitro. Methods RPE cells were co-cultured with THP-1 cells in transwell inserts for 0, 2, 3, 24, 48 and 72 h. Immunofluorescence staining and reverse transcription polymerase chain reaction （RT-PCR） were used to detect the levels of c-fos protein and mRNA in RPE cells. Results RPE cells that were not co-cultured with THP-1 cells revealed either faint or no green fluorescence of c-fos in the cytoplasm and some nuclei. When co-cultured with THP-1 cells for 2 h, green fluorescence labeling of c-fos showed an obvious increase with stronger green fluorescence translocated to the nuclei. After co-culturing with THP-1 cells, c-fos mRNA （0 h, 0.40±0.07） increased rapidly in RPE cells and the enhancement peaked at 2 h （2 h, 0.91±0.10, P=0.002）, and declined to a similar level as the cells that were not co-cultured after 3 h （3 h, 0.51±0.08）. Conclusion Co-culturing with THP-1 cells can quickly up-regulate c-fos protein and mRNA in cultured human RPE cells within a short time period, and induces its nuclear transposition, which suggests the THP-1 cells can activate c-fos.

关 键 词：视网膜色素上皮 THP-1细胞 C-FOS 

分 类 号：R774.1[医药卫生—眼科]