CAJ | 학술논문

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TT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-time PCR)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1 000、2 000 μg/L作用48 h后对TT细胞的增殖抑制率分别为(7.51±1.57)％、(12.76±3.23)％,TT细胞对TRAIL具有明显抵抗性.(2)塞来昔布(celecoxib)联合TRAIL作用48 h,TT细胞凋亡率为(21.92±1.35)％,明显高于单用TRAIL[(4.32±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DR5 mRNA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DR5的表达,促进TRAIL对Caspase-8的裂解,增强TRAIL诱导凋亡的能力.
목적 관찰사망수체4(DR4)급사망수체5(DR5)재종류배사인자상관조망유도배체(TRAIL)유도갑상선수양암TT세포조망중적작용.방법 새서람(MTT)법검측세포증식억제정황;류식세포의검측세포조망;실시정량취합매련반응(Real-time PCR)검측DR4급DR5적기인표체;Western blot검측DR4、DR5급반광안선천동안산특이성단백매(Caspase)-8단백적표체.결과 (1) TRAIL이1 000、2 000 μg/L작용48 h후대TT세포적증식억제솔분별위(7.51±1.57)％、(12.76±3.23)％,TT세포대TRAIL구유명현저항성.(2)새래석포(celecoxib)연합TRAIL작용48 h,TT세포조망솔위(21.92±1.35)％,명현고우단용TRAIL[(4.32±0.83)％]급채용celecoxib[(14.29±2.15)％].(3)단용TRAIL,TT세포DR5단백표체여대조조차이무통계학의의(P＞0.05),차표체량균흔저;이단용celecoxib급celecoxib여TRAIL연합조균능상조TT세포DR5 mRNA급단백적표체,량조차이무통계학의의(P＞0.05),동시연합조능구촉진Caspase-8적렬해,이대DR4적표체칙무명현작용.결론 TT세포대TRAIL적저항성여DR5저표체유관,celecoxib능구상조DR5적표체,촉진TRAIL대Caspase-8적렬해,증강TRAIL유도조망적능력.
Objective To study the role of death receptors death receptor 4 (DR4) and DR5 in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in medullary thyroid cancer TT cell line.Methods The growth inhibition of TT cells was measured by methyl thiazol tetrazolium(MTT) assay.Annexin V/PI double staining was used to analyze the apoptosis rate of TT cells by flow cytometry.The mRNA expression of DR4 and DR5 was detected by using semiquantitative real-time quantitative 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 TT 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 combination 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 expression 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 alone 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 TT cells to TRAIL,which could be reversed by celecoxib through the up-regulation of DR5 and cleavage of Caspase-8.