肿瘤坏死因子相关凋亡诱导配体诱导喉癌凋亡的实验研究

Apoptosis inducing effect by TRAIL on human laryngeal squamous carcinoma Hep-2 cells

目的:研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导喉鳞状细胞癌Hep-2细胞凋亡的作用机制。方法:应用不同浓度的TRAIL作用于喉鳞状细胞癌Hep-2细胞,应用噻唑蓝比色法绘制细胞的生长曲线,计算生长抑制率;应用流式细胞仪检测细胞凋亡率;应用透射电镜观察细胞的微观形态学改变。结果:TRAIL能抑制体外培养的喉癌Hep-2细胞生长,其抑制作用存在着明显的浓度依赖性,随着TRAIL浓度的不断增加,喉癌Hep-2细胞的生长抑制率也增加,经TRAIL(100μg/L)处理24h,喉鳞状细胞癌Hep-2细胞生长抑制率达到(62.75±1.00)%。流式细胞仪检测结果显示喉鳞状细胞癌Hep-2细胞凋亡发生率随TRAIL浓度增加而明显增加,在TRAIL浓度为1、10、100μg/L时凋亡发生率分别为(11.49±0.36)%、(22.31±0.82)%和(59.64±1.10)%,与对照组凋亡发生率为(3.13±0.12)%相比,差异均具有统计学意义(均P<0.01)。透射电镜观察发现,应用100μg/L TRAIL处理后的Hep-2细胞发生凋亡改变,可见细胞出现核碎裂,核仁消失,染色质固缩,电子密度增高,线粒体肿胀,电子密度增大,嵴不清,并有凋亡小体形成。结论:TRAIL能抑制体外培养的喉鳞状细胞癌Hep-2细胞生长,其抑制作用具有明显的浓度依赖性。TRAIL是通过诱导肿瘤细胞凋亡来实现其抑制作用的。

Objective：To study the apoptosis inducing effect of tumor necrosis factor related apoptosis-ligand（TRAIL）on human laryngeal squamous carcinoma Hep-2 cells and its effect mechanism.Method：The human laryngeal squamous carcinoma Hep-2 cell line was treated with different concentration of TRAIL in vitro.The inhibition ratio of tumor cells was determined by MTT colorimetric assay,the incidence of cell apoptosis was determined by flow cytometry method.The morphologic changes of laryngeal squamous carcinoma Hep-2 cell were observed with transmission electron microscope.Result：In vitro,all the different concentrations of TRAIL inhibited laryngeal squamous carcinoma cell＇s growth.The inhibited growth ratio showed significant concentration-dependence.The concentrations for inducing apoptosis-ratio（TRAIL 1,10,100 μg/L）determined by flow cytometry was（11.49±0.36）%,（22.31±0.82）%,（59.64±1.10）% respectively in the study group,and（3.13±0.12）% in the control group,which was significantly different between these two groups（P0.01）.Conclusion：In vitro,TRAIL inhibited the growth of human laryngeal squamous carcinoma Hep-2 cells.The induced apoptosis of TRAIL shows significant concentration-independence.TRAIL inhibits the growth of human laryngeal squamous carcinoma Hep-2 cells trough inducing apoptosis.