摘要
目的探讨高迁移率族蛋白（high mobility group box，HMGB）1对大鼠滑膜细胞STAT信号通路的调控作用。方法将常规培养大鼠滑膜细胞株RSC-364细胞随机分为正常对照组和10μg／LHMGBI刺激组，分别培养6h、12h和24h，RT—PCR检测STAT1／3mRNA的表达，Western印迹法和流式细胞术（flow cytometric analysis，FCM）检测STAT1、STAT3、磷酸化STAT1（phospho—STAT1，p—STAT1）和磷酸化3（phospho—STAT3，p—STAT3）蛋白的表达，免疫细胞化学（ICC）检测PCNA蛋白的表达。结果HMGB1刺激6h～24h组STAT1 mRNA和p-STAT1蛋白的相对表达量呈时间依赖性上调，24h表达最高，与正常对照组相比差异均具有显著统计学意义（P〈0．05，P〈0．01）。PCNA蛋白阳性信号表达于细胞核内，呈棕黄色颗粒，且随着刺激时间延长阳性信号逐渐增强。RT—PCR、ICC染色和FCM均显示STAT3mR—NA和p-STAT3蛋白的相对表达量各组间相比差异均无统计学意义。结论HMGB1可能通过激活STAT1信号转导通路促进滑膜细胞的增殖和分化。
Objective To investigate the effect signal transducer and activator of transcription （STAT） on proliferation process of rat class fibroblast-like synovial cells induced by high mobility group box （HMGB） 1. Methods RSC-364 cells induced by 10μg=/L recombinant HMGB1 were collected in 6 h, 12 h, 24 h respectively, as well as normal control group cells in vitro. The expression of STAT 1 / 3 mRNA were analyzed by RT-PCR technique. Western blotting and flow cytometric analysis （ FCM ） were adopted to detect the expression of STAT1, STAT3, p-STAT1 and p- STAT3. Immunocytoehemistry （ICC） staining was adopted to detect the expression of PCNA protein. Results The relative expression of STAT1 mRNA and p-STAT1 proterin was significantly up-regulated in time-dependent manner from 6 h group to 24 h group, the highest expression of them discovered at 24 h. Compared with normal control group, there were significant statistic differences （P 〈 0. 05 or 0. 01 ） . The positive signal expression of PCNA proteins were localized in nucleus as huffy clumping,and gradually increased in time-dependent manner too. But the changes of STAT3 mRNA and p-STAT3 protein expression indicated no significant statistical differences among normal control group and 6 h - 24 h groups by RT-PCR, ICC staining and FCM. Conclusion HMGB1 could activate STAT1 signal transduct process to promote cell proliferation of rat class fibroblast-like synovial cells.
出处
《医学分子生物学杂志》
CAS
CSCD
2010年第1期33-38,共6页
Journal of Medical Molecular Biology
基金
河北省科技厅科技攻关基金资助项目（No.08206122D）
关键词
高迁移率族蛋白1
滑膜细胞
信号转导和转录激活子
细胞增殖
high mobility group box 1
synoviocytes
signal transducer and activator of transcription
cell proliferation
