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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。
AIM: To investigate the mechanism of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) inducing apoptosis in laryngeal squamous cell carcinoma Hep-2 cells. Methods: Different concentrations of TRAIL were applied to Hep-2 cells in laryngeal squamous cell carcinoma. The cell growth curve was drawn by thiazolyl blue colorimetry, and the growth inhibition rate was calculated. The apoptosis rate was detected by flow cytometry. Transmission electron microscopy Observe the microscopic morphological changes of the cells. RESULTS: TRAIL could inhibit the growth of Hep-2 cells in vitro and there was a significant concentration-dependent inhibition. With the increase of TRAIL concentration, the growth inhibition rate of Hep-2 cells was also increased. After TRAIL (100 μg / L) for 24 h, the growth inhibition rate of Hep-2 cells in laryngeal squamous cell carcinoma reached (62.75 ± 1.00)%. Flow cytometry results showed that the incidence of apoptosis of Hep-2 cells in laryngeal squamous cell carcinoma increased with the increase of TRAIL concentration. The apoptosis rates of Hep-2 cells were (11.49 ± 0.36 ), (22.31 ± 0.82)% and (59.64 ± 1.10)%, respectively, which were significantly different from those in the control group (3.13 ± 0.12)% (all P <0.01). The results of transmission electron microscope showed that apoptotic changes occurred in Hep-2 cells treated with 100 μg / L TRAIL, showing nuclear fragmentation, disappearance of nucleolus, chromatin condensation, electron density increase, mitochondria swelling and electron density increase, Cristae are unclear and apoptotic bodies are formed. CONCLUSION: TRAIL can inhibit the growth of Hep-2 cells cultured in vitro, and its inhibitory effect is obviously concentration-dependent. TRAIL induces tumor cell apoptosis to achieve its inhibitory effect.