白细胞介素-17对人喉癌细胞Hep-2的作用  

Effects of interleukin-17 on human laryngeal carcinoma Hep-2 cells

作　　者：冯舒[1] 王郡甫[2] 陈雪梅[3] 栾俊文[2] 苏庆红[2] 栾萌[2] 许晓群[2] 

机构地区：[1]济南大学山东省医学科学院医学与生命科学学院,250062 [2]山东省医学科学院基础医学研究所 [3]山东大学第二医院耳鼻咽喉头颈外科

出　　处：《国际肿瘤学杂志》2017年第4期241-245,共5页Journal of International Oncology

基　　金：山东省自然科学基金（ZR2013HL028、ZR2014HQ030、ZR2014HM085、ZR2015PC018）;山东省大型科学仪器设备升级改造技术研究专项（2013SJGZ09）;山东省医学科学院医药卫生科技创新工程;山东省中医药科技发展计划（2015-326）

摘　　要：目的 探讨白细胞介素-17（IL-17）对人喉癌细胞Hep-2增殖、凋亡和迁移的作用.方法将IL-17瞬时转染Hep-2细胞,即转染IL-17组,同时设置空载体组（pEGFP-N1）和正常对照组.荧光显微镜观察转染情况,反转录-聚合酶链反应（RT-PCR）和Western blotting分别检测转染前后IL-17 mRNA和蛋白的表达,四甲基偶氮唑蓝（MTT）法检测转染前后细胞增殖能力变化,流式细胞术检测转染前后细胞凋亡变化,细胞划痕修复实验和Transwell小室检测转染前后细胞迁移能力变化.结果 荧光显微镜下可以看到转染空载体pEGFP-N1和转染目的基因IL-17的Hep-2细胞出现绿色荧光.成功转染IL-17后Hep-2细胞在mRNA和蛋白水平均有IL-17的表达.与正常对照组相比,转染48 h后,转染IL-17组细胞的增殖能力明显减弱（0.34±0.03∶0.46±0.04,P=0.006）.转染IL-17组细胞凋亡率明显高于正常对照组（26.80%±0.80%∶2.90%±0.31%,P=0.000）.细胞划痕修复实验结果显示,转染IL-17组细胞相对划痕宽度明显大于正常对照组（1.59±0.01∶1.36±0.01,P=0.000）.Transwell小室迁移实验结果显示,转染IL-17组细胞明显低于正常对照组细胞的迁移数量（26.33±2.08∶49.33±1.53,P=0.000）.结论 IL-17能够抑制人喉癌细胞Hep-2的增殖能力,降低其迁移能力,增强其凋亡.因此,IL-17可通过多种途径抑制喉癌的发生发展.Objective To investigate the effects of interleukin-17 （IL-17） on the cell proliferation, apoptosis and migration of human laryngeal carcinoma Hep-2 cells.Methods IL-17 was transiently transfected into Hep-2 cells, and at the same time empty vector group （pEGFP-N1） and normal control group were set up.The efficiency of transfection was evaluated by fluorescence microscope, and the mRNA and protein expressions of IL-17 were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting.The proliferation of cells was detected by methyl thiazolyl tetrazolium （MTT） method, and the apoptosis was detected by flow cytometry.The migration ability was detected by wound-healing assay and Transwell assay.ResultsHep-2 cells transfected with empty vector pEGFP-N1 and IL-17 showed green fluorescence under the fluorescence microscope.Hep-2 cells expressed IL-17 at both mRNA and protein levels after transfection with IL-17.Compared with the normal control group, the proliferation of IL-17 transfected Hep-2 cells was significantly inhibited after 48 h transfection （0.34±0.03 vs.0.46±0.04, P=0.006）.The apoptotic rate of IL-17 transfected cells was higher than that of normal control group （26.80%±0.80% vs.2.90%±0.31%, P=0.000）.According to the wound-healing assay, compared with the normal control group, the scratch width of IL-17 transfected cells was significantly greater （1.59±0.01 vs.1.36±0.01, P=0.000）.Transwell migration experiment showed that the migration of IL-17 transfected cells was significantly lower than that of the normal control group （26.33±2.08 vs.49.33±1.53, P=0.000）.Conclusion IL-17 can inhibit the proliferation of human laryngeal carcinoma Hep-2 cells, reduce their migration ability and enhance their apoptosis ability.Therefore, IL-17 may inhibit the occurrence and development of laryngeal carcinoma through a variety of mechanisms.

关 键 词：白细胞介素17 喉肿瘤 细胞增殖 细胞凋亡 细胞运动 

分 类 号：R739.65[医药卫生—肿瘤]