中华内分泌外科杂志
中華內分泌外科雜誌
중화내분비외과잡지
CHINESE JOURNAL OF ENDOCRINE SURGERY
2014年
5期
398-401
,共4页
郑桂彬%孟宪瑛%韩佳滨%张强%杨帅
鄭桂彬%孟憲瑛%韓佳濱%張彊%楊帥
정계빈%맹헌영%한가빈%장강%양수
肿瘤坏死因子相关凋亡诱导配体%甲状腺髓样癌%自噬%凋亡%Beclin 1
腫瘤壞死因子相關凋亡誘導配體%甲狀腺髓樣癌%自噬%凋亡%Beclin 1
종류배사인자상관조망유도배체%갑상선수양암%자서%조망%Beclin 1
TRAIL%Medullary thyroid cancer%Autophagy%Apoptosis%Beclin1
目的 观察在肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)作用下甲状腺髓样癌TT细胞自噬发生情况,明确自噬在TRAIL诱导甲状腺癌TT细胞凋亡中的作用.方法 采用MTT法检测细胞增殖抑制率;MDC染色检测自噬的发生情况;流式细胞仪检测细胞凋亡;Western blot检测相关蛋白的表达.结果 ①MTT实验显示:TRAIL对TT细胞增殖抑制作用弱,在浓度为250、500、1000、2000 ng/ml的TRAIL作用48 h后对TT细胞增殖抑制率分别为(3.02±1.82)％、(4.87±1.45)％、(7.51±1.57)％及(12.76±3.23)％;②TRAIL作用48 h后,胞内代表自噬小体的绿色荧光亮点明显增多,且随TRAIL浓度的增加而增加;③3-MA预处理TT细胞4h,TRAIL 500 ng/ml及1000 ng/ml的凋亡率分别为(17.83 ±1.54)％及(27.81±1.79)％,明显高于单用TRAIL(3.70±0.34、6.55 ±0.59)％与3-MA (7.71±0.64)％之和,差异有统计学意义(t=3.282,P＜0.05;t=7.830,P＜0.01).④各实验组可见明显的caspase-8的活化裂解片段,自噬相关蛋白Beclin 1的表达明显减低.结论 甲状腺髓样癌TT细胞对TRAIL不敏感,TRAIL能诱导TT细胞内自噬发生,抑制自噬可明显改善TT细胞对TRAIL的敏感性.
目的 觀察在腫瘤壞死因子相關凋亡誘導配體(tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)作用下甲狀腺髓樣癌TT細胞自噬髮生情況,明確自噬在TRAIL誘導甲狀腺癌TT細胞凋亡中的作用.方法 採用MTT法檢測細胞增殖抑製率;MDC染色檢測自噬的髮生情況;流式細胞儀檢測細胞凋亡;Western blot檢測相關蛋白的錶達.結果 ①MTT實驗顯示:TRAIL對TT細胞增殖抑製作用弱,在濃度為250、500、1000、2000 ng/ml的TRAIL作用48 h後對TT細胞增殖抑製率分彆為(3.02±1.82)％、(4.87±1.45)％、(7.51±1.57)％及(12.76±3.23)％;②TRAIL作用48 h後,胞內代錶自噬小體的綠色熒光亮點明顯增多,且隨TRAIL濃度的增加而增加;③3-MA預處理TT細胞4h,TRAIL 500 ng/ml及1000 ng/ml的凋亡率分彆為(17.83 ±1.54)％及(27.81±1.79)％,明顯高于單用TRAIL(3.70±0.34、6.55 ±0.59)％與3-MA (7.71±0.64)％之和,差異有統計學意義(t=3.282,P＜0.05;t=7.830,P＜0.01).④各實驗組可見明顯的caspase-8的活化裂解片段,自噬相關蛋白Beclin 1的錶達明顯減低.結論 甲狀腺髓樣癌TT細胞對TRAIL不敏感,TRAIL能誘導TT細胞內自噬髮生,抑製自噬可明顯改善TT細胞對TRAIL的敏感性.
목적 관찰재종류배사인자상관조망유도배체(tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)작용하갑상선수양암TT세포자서발생정황,명학자서재TRAIL유도갑상선암TT세포조망중적작용.방법 채용MTT법검측세포증식억제솔;MDC염색검측자서적발생정황;류식세포의검측세포조망;Western blot검측상관단백적표체.결과 ①MTT실험현시:TRAIL대TT세포증식억제작용약,재농도위250、500、1000、2000 ng/ml적TRAIL작용48 h후대TT세포증식억제솔분별위(3.02±1.82)％、(4.87±1.45)％、(7.51±1.57)％급(12.76±3.23)％;②TRAIL작용48 h후,포내대표자서소체적록색형광량점명현증다,차수TRAIL농도적증가이증가;③3-MA예처리TT세포4h,TRAIL 500 ng/ml급1000 ng/ml적조망솔분별위(17.83 ±1.54)％급(27.81±1.79)％,명현고우단용TRAIL(3.70±0.34、6.55 ±0.59)％여3-MA (7.71±0.64)％지화,차이유통계학의의(t=3.282,P＜0.05;t=7.830,P＜0.01).④각실험조가견명현적caspase-8적활화렬해편단,자서상관단백Beclin 1적표체명현감저.결론 갑상선수양암TT세포대TRAIL불민감,TRAIL능유도TT세포내자서발생,억제자서가명현개선TT세포대TRAIL적민감성.
Objective To observe the level of autophagy induced by TRAIL in TT cell line and identify the role of autophagy in TRAIL-inducing apoptosis of TT cell line.Methods The growth inhibition of TT cells was measured by MTT assay.MDC staining was used to identify the happening of autophagy.Annexin V/PI double staining was used to analyze the apoptosis rate of TT cells by flowcytometry.The protein expression of caspase-8 and Beclin1 was detected by Western blot.Results ① The growth inhibition ratio of TT cells induced by TRAIL at the concentration of 250,500,1000 and 2000 ng/ml was (3.02 ± 1.82)％,(4.87 ± 1.45)％,(7.51 ± 1.57) ％,(12.76 ± 3.23) ％ respectively,which suggested significant resistance of TT cells to TRAIL.② MDC-labeled green light vesicles was significantly increased after the treatment of TRAIL for 48 h.③ The apoptosis rate of TT cells induced by TRAIL at the concentration of 500 ng/ml and 1000 ng/ml after the pretreat ment of 3-MA for 4 h was(17.83 ± 1.54) ％ and(27.81 ± 1.79) ％ respectively,which was significantly higher than the apoptosis rate induced by TRAIL(3.70 ± 0.34) ％,(6.55 ± 0.59) ％ alone and that induced by 3-MA(7.71 ± 0.64) ％ (t =3.282,P ＜ 0.05 ; t =7.830,P ＜ 0.01).④ The combination treatment of TRAIL and 3-MA increased the cleavage of caspase-8 and down-regulated the expression of Beclin 1.Conclusion Autophagy induced by TRAIL may contributes to the resistance of TT cells to TRAIL,which can be reversed by the inhibition of autophagy.
