白细胞介素-37在人HepG2细胞中的表达  

Expression of interleukin-37in human HepG2cells

作　　者：刘航[1,2] 李为民[1] 刘浩润[1] 赵洪强[1] 王瑞官[1] 龙婷 LIU Hang LI Wei-min LIU Hao-run ZHAO Hong-qiang WANG Rui-guan LONG Ting(Department of Liver and Gallbladder Surgery,the People＇s Liberation Army 309 Hospital, Beijing 100091 China Beijing Huairou Hospital,Beijing Huairou 101400,China)

机构地区：[1]解放军第309医院肝胆外科,北京100091 [2]北京怀柔医院,北京市怀柔区101400

出　　处：《临床消化病杂志》2016年第6期354-357,共4页Chinese Journal of Clinical Gastroenterology

摘　　要：[目的]研究白细胞介素-37(interleukin-37,IL-37)在人肝肿瘤细胞系(HepG2)细胞中的表达,及在炎症因子刺激下,HepG2细胞中IL-37表达水平的改变。[方法]在37℃、5%CO2条件下培养人HepG2细胞,以脂多糖(lipopolysaccharide,LPS)刺激细胞,分别于刺激24h、48h、72h后提取细胞,分别提取细胞总RNA、总蛋白,利用逆转录酶聚合链反应技术(RT-PCR)检测细胞中IL-37-mRNA的表达,利用Western blot技术检测细胞中IL-37蛋白的表达水平。并且在LPS刺激细胞72h后,运用免疫荧光和激光共聚焦技术检测IL-37在细胞中的表达定位。[结果]RT-PCR结果证明HepG2中存在IL-37-mRNA的表达,且随着LPS刺激时间的延长IL-37-mRNA表达增多,0h(无刺激)、24h、48h、72h灰度值分别为0.10±0.008、0.14±0.014、0.22±0.010、0.28±0.014;Western blot结果显示细胞中存在IL-37蛋白的表达,且其表达趋势与IL-37mRNA表达趋势相同,即随着刺激时间延长,HepG2中IL-37蛋白表达增多,由24h、48h、72h分别为0.54±0.16、0.87±0.04、1.51±0.18;免疫荧光共聚焦结果显示IL-37在细胞核和细胞质中均有表达,以细胞核周围表达最为丰富。[结论]人HepG2细胞中存在IL-37表达,炎症因子刺激后,IL-37mRNA及IL-37蛋白表达水平均呈升高,趋势相同;IL-37在HepG2细胞质内和细胞核内均表达,以细胞核周围表达最为丰富。[Objective]To investigate the expression level of Interlukin-37（IL-37）in human hepatoeellu- lar liver carcinoma cell line（HepG2）and its change induced by lipopolysaecharide（LPS）. [Methods]HepG2 cells were cultured in a suitable environment（37℃ ,5% CO2 ）,stimulated by with LPS,extract the cells at the time of 24 h,48 h,72 h after the stimulation. The total RNA and protein of the cell were extracted re- spectively. By using the method of RT-PCR and Western blot,the expression levels of IL-37 mRNA and IL- 37 protein were determined. At the time point of 72 h after the stimulation,the disposition of the IL-37 in- side the cell was observed by using the immunofluorescence laser scanning confoeal microscope. [Results] The RT-PCR demonstrated the expression of the II.-37 mRNA in HepG2. After the stimulation of the LPS,the intensity of IL-37 mRNA from 0 h to 72 h were 0.10±0. 008,0.14±0. 014,0. 22±0. 010,0.28± 0. 014 respectively,the similar increased trend was observed in IL-37 protein expression. The immunofluo- rescence microscope found that the IL-37 expressed both in the nucleus and the cytoplasm, especially a- round the nucleus. [Conclusion]IL-37 can positively expressed in normal HepG2 cells. The stimulation with the LPS can increase the expression level of IL-37 mRNA and IL-37 protein. IL-37 can be observed both in the nucleus and the cytoplasm,especially around the nucleus.

关 键 词：肝肿瘤细胞系 白细胞介素-37 脂多糖 表达水平 

分 类 号：R735.7[医药卫生—肿瘤]