鞭毛蛋白活化TLR5信号途径对于SPC-A-1细胞生物学行为的影响  

Influence of activation of TLR5 signaling pathway stimulated by Flagellin on the biological action of SPC-A-1 cells

作　　者：周辉 马安迪 文亦戈 李芳 罗媚 罗永忠 易青 陈建华 

机构地区：[1]湖南省肿瘤医院/中南大学湘雅医学院附属肿瘤医院胸部内一科,长沙410013

出　　处：《中国医师杂志》2015年第10期1485-1490,共6页Journal of Chinese Physician

基　　金：基金项目：湖南省自然科学基金资助项目（2015JJ6058）;湖南省卫计委资助项目（B2015-111）

摘　　要：目的 探讨外源性配体鞭毛蛋白活化TLR5后,对于SPC-A-1细胞生物学行为的影响.方法 选用SPC-A-1细胞作为研究对象,鞭毛蛋白作用浓度为0.1μg/ml.构建TLR5-shRNA,并转染SPC-A-1细胞.实验分为四组：对照组、TLR5-shRNA组、Flagellin组和TLR5-shRNA＋ Flagellin组.用四甲基偶氮唑蓝比色法（MTT）在0、24、48、72 h检测细胞增殖情况,在48 h分别用TUNEL法检测细胞凋亡,Transwell细胞侵袭实验研究细胞侵袭能力的影响,平板划线培养实验研究细胞迁移能力的影响,细胞集落形成实验研究细胞克隆形成能力.结果 （1）于0、24、48、72 h在570 nm波长处检测各组细胞的光密度值,结果显示,在同一时间点,Flagellin组与其它三组相比,细胞增殖明显受到抑制（P＜0.01）,TLR5-shRNA＋ Flagellin组与对照组和TLR5-shRNA组相比,细胞增殖无明显影响（P＞0.05）.（2）对照组、TLR5-shRNA组、Flagellin组和TLR5-shRNA＋ Flagellin组的细胞凋亡率分别为（14.75 ±0.072）％、（15.09 ±0.053）％、（21.63 ±0.047）％、（14.98 ±0.036）％.与对照组、TLR5-shRNA组和TLR5-shRNA＋ Flagellin组相比,Flagellin组细胞凋亡率明显增加（P＜0.05）.（3）对照组、TLR5-shRNA组、Flagellin组和TLR5-shRNA＋ Flagellin组穿膜细胞数分别为39.55±2.31、42.78±3.52、18.75±3.77和38.43±2.98.与对照组、TLR5-shRNA组和TLR5-shRNA＋ Flagellin组相比,Flagellin组的细胞其侵袭能力明显受到抑制（P＜0.01）,而对照组、TLR5-shRNA组和TLR5-shRNA＋ Flagellin组两两比较,差异均无统计学意义（P＞0.05）.（4）与对照组、TLR5-shRNA组和TLR5-shRNA＋ Flagellin组相比,Flagellin刺激48 h后,Flagellin组细胞的迁移能力显著下降（P＜0.05）,而与对照组、TLR5-shRNA组相比,TLR5-shRNA＋ Flagellin组细胞的迁移能力无明显改变（P＞0.05）.（5）对照组、TLR5-shRNA组、Flagellin组和TLR5-shRNA＋ Flagellin组细胞集落形成率分别为（40.37±3.88）％、（39.88±2.49�Objective To investigate the influence of the activation of Toll-like receptors （TLR） 5 signaling pathway stimulated by flagellin on the biological action of SPC-A-1 cells.Methods SPC-A-1 cells were chosen as the research object, and the concentration of flagellin was 0.1 μg/ml.Four groups were control, TLR5-shRNA, Flagellin, and TLR5-shRNA ＋ Flagellin groups.Cell proliferation was detected with methyl thiazolyl tetrazolium （MTT） at 0 h, 24 h, 48 h and 72 h, cell apoptosis with TdT-mediated dUTP nick-end labeling （TUNEL） assay, cell invasion with Transwell cell invasion assay, cell migration with impact streak plate culture experiments, and cell colony formation with cell colony formation experimental at 48 h.Results （1）At 0 h, 24 h, 48 h and 72 h, the optical density values was detected under 570 nm wavelength.At the same time point, compared to the other groups, the cell proliferation of the Flagellin group was significantly inhibited （P 〈 0.01）.Compared to control and TLR5-shRNA groups,there was no significant effect in cell proliferation of TLR5-shRNA ＋ Flagellin group （P 〉 0.05）.（2） The apoptosis rates of the control, TLR5-shRNA, Flagellin, and TLR5-shRNA ＋ Flagellin groups were （14.75 ±0.072）%, （15.09 ±0.053）%, （21.63% ±0.047）% and （14.98 ±0.036）%, respectively.Compared to the other groups, the apoptosis rate of the Flagellin group was significantly increased （P 〈 0.05）.（3）The number of penetrating cells of the control, TLR5-shRNA, Flagellin, and TLR5-shRNA ＋ Flagellin was 39.55 ± 2.31, 42.78 ± 3.52, 18.75 ± 3.77 and 38.43 ± 2.98, respectively.Compared to the other groups, cell invasion of the Flagellin group was significantly inhibited （P 〈 0.01） , while pairwise comparisons of the control, TLR5-shRNA, and TLR5-shRNA ＋ Flagellin groups, there was significant difference （P 〉 0.05）.（4）Compared to the other groups, cell migration of Flagellin group was significantly declined after 48h stimulated by Flagellin （P 〈0

关 键 词：Toll样受体5/药物作用 鞭毛蛋白/药理学 信号传导 肺肿瘤/药物疗法 

分 类 号：R734.2[医药卫生—肿瘤]