摘要
目的 深入研究维甲酸诱导基因G（RIG-G）表达中干扰素α（IFN-α）和全反式维甲酸（ATRA）两条信号途径间的相互关系.方法 运用急性早幼粒细胞白血病细胞系NB4细胞和信号转导子与转录激活子（STAT）1缺失的人纤维肉瘤细胞系U3A细胞,检测ATRA作用于NB4细胞后,细胞内STAT2的磷酸化水平,并测定NB4细胞培养液上清及转染了干扰素调节因子1（IRF-1）的U3A细胞培养液上清中的IFN-α水平,以及培养液上清对STAT2的磷酸化和RIG-G基因表达的诱导作用.结果 ATRA处理72 h后,NB4细胞内的STAT2磷酸化水平和细胞培养液上清中的IFN-α浓度均明显升高[ IFN-α：（ 7.6±0.3）pg/ml比（1.5±0.5）pg/ml,P＜0.05].该培养液上清能够诱导STAT2发生磷酸化,并上调RIG-G的蛋白水平.U3A细胞转染IRF-1后,培养液上清中的IFN-α浓度也明显提高[（8.8±1.4） pg/ml比（3.4±0.4）pg/ml,P＜0.05].结论 RIG-G基因的表达与ATRA和IFN-α这两条信号途径的协同作用密切相关,ATRA可以通过上调IRF-1的蛋白水平促进细胞分泌IFN-α,所分泌的IFN-α能诱导STAT2发生磷酸化,并进一步增强细胞内RIG-G基因的表达.
Objective To explore the relationship between interferon （IFN） α and all-trans retinoic acid （ATRA）-induced signaling pathways in the expression of retinoic acid-induced gene G （RIG-G）.Methods Acute promyelocytic leukemia cell line NB4 and signal transducer and activator of transcription （STAT） 1-deficient U3A cells were used. The protein levels of tyrosine-phosphorylated STAT2 in ATRA-treated NB4 cells were detected by Western blot.The culture supernatants of NB4 cells treated with ATRA for different time or U3A cells transfected with interferon regulatory factor （IRF）-1 were respectively collected. And the concentration of IFN-α was determined by enzyme-linked immunosorbent assay （ELISA）.The effects of NB4 cell culture supernatants on the phosphorylation of STAT2 and the expression of RIG-G were detected by Western blot.Results The level of phosphorylated-STAT2 was obviously upregulated in NB4 cells treated with ATRA for 72 hour,as well as the concentration of IFN-α in culture supernatants. The concentration of IFN-α increased from （1.5 ± 0.5 ） pg/ml in the untreated group to （7.6 ±0.3） pg/ml （P 〈0.05）.After a 96-hour treatment,the concentration of IFN-α was up to （63.8 ±5.8） pg/ml. And these culture supernatants could induce the tyrosine phosphorylaton of STAT2 and upregulate the protein level of RIG-G. As for U3A cells transfected with IRF-1,the concentration of IFN-αfrom the culture supernatant also increased 3-fold versus the control group transfected with empty vectors [（8.8±1.4） pg/ml vs （3.4 ±0.4）pg/ml,P〈0.05].Conclusions RIG-G gene expression is closely correlated with the cross-talk between ATRA and IFN-α -induced signaling pathways.ATRA increases the secretion of IFN-α by up-regulating the protein level of IRF-1.Then the secreted IFN-α may induce the phosphorylation of STAT2 and reinforce the expression of RIG-G.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第2期124-127,共4页
National Medical Journal of China
基金
基金项目：国家自然科学基金（30971275）
上海市领军人才“困家队”专项基金
上海市科委优秀学术带头人计划项目（11XD1403500）
关键词
白血病
早幼粒细胞
急性
维甲酸
干扰素Α
维甲酸诱导基因G
信号转导
Leukemia, promyelocytic, acute
Retinoic acid
Interferon-alpha
Retinoic acidinduced gene G
Signaling pathway
