干扰素α对口腔鳞状细胞癌细胞JAK-信号转导子和转录激活子-细胞因子信号抑制因子信号通路基因表达的影响  

Interferon alpha on oral squamons cell cancer JAK signal transduction and transcriptional activation suppressor cytokine signal pathways of gene expression

作　　者：游锦梅[1] 许雅鑫[2] 吴彬 Yang Yong 李钰[5] 陈显久[1] 

机构地区：[1]山西医科大学生物化学与分子生物学教研室,太原030001 [2]山西医科大学流行病学教研室,太原030001 [3]太原钢铁集团有限公司总医院中心实验室 [4]Department of Epidemiology, Medical Board, Singapore General Hospital [5]山西医科大学第一医院骨科

出　　处：《肿瘤研究与临床》2015年第5期320-327,331,共9页Cancer Research and Clinic

基　　金：山西省国际科技合作项目（2012081050-1）;山西医科大学创新项目（01201002）

摘　　要：目的 以3种不同癌变程度的口腔细胞为工具,观察在干扰素α（IFN-α）作用下,JAK-信号转导子和转录激活子（STAT）-细胞因子信号抑制因子（SOCS）信号分子的表达变化,为深入理解口腔鳞状细胞癌（OSCC）细胞免疫逃逸的机制提供研究基础.方法 常规培养NOK、DOK、KB细胞.空白组加入完全培养液,二甲基亚砜（DMSO）对照组加入含有0.1％ DMSO的完全培养液,实验组设10、100、500U/ml三个不同浓度组,每孔分别加入含不同浓度IFN-α的完全培养液,JAK抑制剂干预组在加入IFN-α前1h加入JAK抑制剂CP-690550 100 μmol/L.细胞培养24 h后检测.进行反转录聚合酶链反应（RT-PCR）和Western blot实验.结果 采用RT-PCR法检测显示：JAK1和JAK2在NOK细胞均有少量表达,IFN-α和CP-690550对NOK细胞组的JAK1和JAK2表达无影响,差异无统计学意义（P＞0.05）.100和500 U/ml的IFN-α能刺激DOK和KB细胞的JAK1和JAK2表达增加,差异均具有统计学意义（P＜0.05）.CP-690550能有效减低DOK和KB细胞的JAK1表达,差异均具有统计学意义（P＜0.05）,而对JAK2表达无作用.采用Western blot法检测显示：STAT1、STAT3和pSTAT3（Tyr705）在对照组均有表达,pSTAT1（Tyr701）在对照组无表达;IFN-α和CP-690550对NOK细胞组的STAT1、STAT3和pSTAT3（Tyr705）表达无影响,差异无统计学意义（P＞0.05）.100、500 U/ml的IFN-α能刺激DOK和KB细胞的pSTAT3（Tyr705）表达增加,差异均具有统计学意义（P＜0.05）;而对pSTAT1（Tyr701）的表达无影响.CP-690550能有效减低DOK和KB细胞的pSTAT3（Tyr705）表达,差异均具有统计学意义（P＜0.05）.采用Western blot法检测显示,SOCS1和SOCS3在对照组均有表达;IFN-α和CP-690550对NOK细胞组的SOCS1和SOCS3表达无影响,差异无统计学意义（P＞0.05）.100 U/ml和500 U/ml的IFN-α能刺激DOK和KB细胞的SOCS1表达增加,差异均具有统计学意义（P＜0.05）;而对SOCS3的表达无影响.CP-690550能有效减低DOObjective To observe the change of JAK-STAT-SOCS signal molecules after interferons alpha acting on the cancerous oral cells in 3 different degrees,namely NOK,DOK and KB cells,and to provide research foundation for the deep understanding of OSCC （oral squamous cell cancer） tumor ceils immune escape mechanism.Methods NOK,DOK,and KB cells were all cultured respectively,and then the third passage cells in the logarithmic growth phase were inoculated in cell culture plate.Blank control group of each hole was added 2 ml complete medium containing 10 % FPS.DMSO control group of each hole was added 2 ml complete medium containing 0.1% DMSO.And in experimental groups containing 10 U/ml,100 U/ml,and 500 U/ml interferons,complete culture medium were added to each hole containing different concentrations of interferons alpha.CP-690550 （100 μmol/L） was added before interferons alpha was added 1 h.All were detected by RT-PCR test and Western blot test after conventional cultured for 24 h.Results RT-PCR detection showed that JAK1 and JAK2 in NOK cells had a small amount of expression,interferons alpha and CP-690550 cells could not influence the expression of JAK1 and JAK2 of NOK group,and there was no statistically significant difference （P 〉 0.05）.Interferons alpha in 100 and 500 U/ml could stimulate the increase of JAK1 and JAK2 expression in DOK and KB cells,and the differences were statistically significant （P 〈 0.05）.CP-690550 could effectively reduce the JAK1 expression of DOK and KB cells,while had no effect on the expression of JAK2,and the differences were statistically significant （P 〈 0.05）.Western blot showed that STAT1,STAT3 and pSTAT3 （Tyr705） all expressed in the control group,while pSTAT1 （Tyr701） didn＇t express in the control group.Interferons alpha and CP-690550 cells had no effect on STAT1,STAT3 and pSTAT3 （Tyr705） expression of NOK group,and there was no statistically significant difference （P 〉 0.05）.100 U/ml and 500 U/ml of interferons alpha could stimulate t

关 键 词：干扰素 口腔鳞状细胞癌 酪氨酸蛋白激酶 信号转导通路 

分 类 号：R739.8[医药卫生—肿瘤]