摘要
目的观察死亡受体4（DR4）及死亡受体5（DR5）在肿瘤坏死因子相关凋亡诱导配体（TRAIL）诱导甲状腺髓样癌r丌细胞凋亡中的作用。方法噻唑蓝（MTT）法检测细胞增殖抑制情况；流式细胞仪检测细胞凋亡；实时定量聚合酶链反应（Real-timePCR）检测DR4及DR5的基因表达；Western blot检测DR4、DR5及半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-8蛋白的表达。结果（1）TRAIL以1000、2000μg/L作用48h后对TT细胞的增殖抑制率分别为（7．51±1．57）％、（12．76±3．23）％，TF细胞对TRAIL具有明显抵抗性。（2）塞来昔布（celecoxib）联合TRAIL作用48h，TT细胞凋亡率为（21．924±1．35）％，明显高于单用TRAIL[（4．324±0．83）％]及采用celecoxib[（14．29±2．15）％]。（3）单用TRAIL，TT细胞DR5蛋白表达与对照组差异无统计学意义（P〉0．05），且表达量均很低；而单用celecoxib及celecoxib与TRAIL联合组均能上调TT细胞DR5mRNA及蛋白的表达，两组差异无统计学意义（P〉0．05），同时联合组能够促进Caspase-8的裂解，而对DR4的表达则无明显作用。结论TT细胞对TRAIL的抵抗性与DR5低表达有关，celecoxib能够上调DR．5的表达，促进TRAIL对Caspase-8的裂解，增强TRAIL诱导凋亡的能力。
Objective To study the role of death receptors death receptor 4 （DR4） and DR5 in tumor necrosis factor-related apoptosisinducing ligand （TRAIL）-induced apoptosis in medullary thyroid cancer TF cell line. Methods The growth inhibition of 3T cells was measured by methyl thiazol tetrazoli- um（MTT） assay. Annexin V/PI double staining was used to analyze the apoptosis rate of 3T cells by flow cytometry. The mRNA expression of DR4 and DR5 was detected by using semiquantitative real-time quanti tative polymerase chain reaction （Real-time PCR）. The protein expression of DR4, DR5 and Caspase-8 was detected by using Western blotting. Results （ 1 ） The growth inhibition ratio of 3T cells induced by TRAIL at the concentration of 1 000 μg/L and 2 000 μg/L was （7.51 ± 1.57） % and （ 12. 76 ±3.23 ） % respectively, suggesting a significant resistance of TT cells to TRAIL; （2） The apoptosis rate of combina tion group after treatment for 48 h was （21.92 ＋ 1.35 ）%, which was significantly higher than TRAIL [ （4.32 ± 0. 83） % ] and celecoxib [ （ 14. 29 ± 2. 15 ） % ] treatment alone （ P 〈 0. 01 ） ; （3） The expres sion of DR5 was low in control group and TRAIL group, but there was no significant difference between the two groups （P 〉 0. 05）. The co-administration of TRAIL and celecoxib and administration of celecoxib a lone could up-regulate the transcripts and translation of DR5 （ P 〉 0. 05 ）, but not DR4. The cleavage of Caspase-8 could be augmented by the combined treatment of TRAIL and celecoxib. Conclusion The low expression of DR5 was involved in the resistance of TF cells to TRAIL, which could be reversed by celecox ib through the up-regulation of DR5 and cleavage of Caspase-8.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第4期750-752,共3页
Chinese Journal of Experimental Surgery
基金
古林省科委国际合作项目（20110136）
关键词
死亡受体
肿瘤坏死因子相关凋亡诱导配体
塞来昔布
甲状腺髓样癌
Death receptor
Tumor necrosis factor-related apoptosis-inducing ligand
Celecoxib
Medullary thyroid cancer
作者简介
孟宪瑛，Email：xy6823@163．com
