CAJ | 학술논문

目的探讨甲状腺激素促进肿瘤细胞增殖及血管新生的信号通路.方法体外培养人胶质母细胞瘤细胞系(SNB19),给予甲状腺激素(主要为T4,100 nmol/L)、四碘甲腺乙酸(tetraiodothyroacetic acid,Tetrac,100 nmol/L)、蛋白激酶C(PKC)抑制剂(2.5 μmol/L)作用后,采用Western印迹方法检测磷酸化蛋白激酶D1(PKD1)、磷酸化组蛋白去乙酰化酶(HDAC)5、磷酸化细胞外信号调节激酶(ERK)1/2的表达,ELISA方法检测细胞培养上清血管内皮生长因子(VEGF)的表达量,3-(4,5)-2-唑噻-(2,5)-二苯基溴化四氮唑蓝(MTT)比色法检测细胞增殖.结果与对照组相比,T4干预组磷酸化PKD1、磷酸化HDAC5、磷酸化ERK1/2水平均增加(P均＜0.05),Tetrac干预组及PKC抑制剂干预组磷酸化PKD1、磷酸化HDAC5、磷酸化ERK1/2水平均降低(P均＜0.05).ELISA结果显示,与对照组相比,T4干预组VEGF浓度升高[(56.763±2.611) ng/L vs.(36.597±0.933) ng/L,P＜0.05],Tetrac+T4干预组VEGF浓度降低[(22.215±1.531) ng/L vs.(36.597±0.933) ng/L,P＜0.05].MTT结果显示,与对照组相比,T4干预组OD值较高[(0.333±0.020)vs.(0.243±0.006),P＜0.05],Tetrac干预组OD值较低[(0.060±0.016) vs.(0.243±0.006),P＜0.05].结论甲状腺激素通过结合整合素αvβ3,激活ERK1/2信号通路促进肿瘤细胞增殖,激活PKC/PKD1/HDAC5信号通路促进血管新生.
목적 탐토갑상선격소촉진종류세포증식급혈관신생적신호통로.방법 체외배양인효질모세포류세포계(SNB19),급여갑상선격소(주요위T4,100 nmol/L)、사전갑선을산(tetraiodothyroacetic acid,Tetrac,100 nmol/L)、단백격매C(PKC)억제제(2.5 μmol/L)작용후,채용Western인적방법검측린산화단백격매D1(PKD1)、린산화조단백거을선화매(HDAC)5、린산화세포외신호조절격매(ERK)1/2적표체,ELISA방법검측세포배양상청혈관내피생장인자(VEGF)적표체량,3-(4,5)-2-서새-(2,5)-이분기추화사담서람(MTT)비색법검측세포증식.결과 여대조조상비,T4간예조린산화PKD1、린산화HDAC5、린산화ERK1/2수평균증가(P균＜0.05),Tetrac간예조급PKC억제제간예조린산화PKD1、린산화HDAC5、린산화ERK1/2수평균강저(P균＜0.05).ELISA결과현시,여대조조상비,T4간예조VEGF농도승고[(56.763±2.611) ng/L vs.(36.597±0.933) ng/L,P＜0.05],Tetrac+T4간예조VEGF농도강저[(22.215±1.531) ng/L vs.(36.597±0.933) ng/L,P＜0.05].MTT결과현시,여대조조상비,T4간예조OD치교고[(0.333±0.020)vs.(0.243±0.006),P＜0.05],Tetrac간예조OD치교저[(0.060±0.016) vs.(0.243±0.006),P＜0.05].결론 갑상선격소통과결합정합소αvβ3,격활ERK1/2신호통로촉진종류세포증식,격활PKC/PKD1/HDAC5신호통로촉진혈관신생.
Objective To study the signaling pathway of proliferation and angiogenesis of tumor cells promoted by thyroid hormone.Methods Human glioblastoma cells(SNB 19) were cultured with thyroid hormone (mainly T4,100 nmol/L),tetraiodothyroacetic acid (Tetrac,100 nmol/L) or protein kinase C (PKC) inhibitor (2.5 μmol/L)in vitro.The expression of phosphorylated protein kinase D 1 (PKD1),phosphorylated histone deacetylase (HDAC)5 and phosphorylated extracellular regulated protein kinases (ERK)1/2 were detected by Western blots.The expression of vascular endothelial growth factor (VEGF) in supernatant was measured by ELISA.The proliferation of SNB19 cells was detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) colorimetric method.Results Compared with control group,phosphorylated PKD1,phosphorylated HDAC5 and phosphorylated ERK1/2 were all increased in T4 group (all P＜0.05),and decreased in Tetrac+T4 group and PKC inhibitor group (all P ＜0.05).The concentration of VEGF in T4 group was enhanced [(56.763 ± 2.611) ng/L vs.(36.597 ± 0.933) ng/L,P ＜0.05],while reduced in Tetrac+T4 group [(22.215 ± 1.531) ng/L vs.(36.597 ± 0.933) ng/L,P ＜0.05],compared with control group.The results of MTT showed that compared with control group,the OD value of T4 group was enhanced[(0.333 ± 0.020) vs.(0.243 ± 0.006),P＜0.05],and reduced in Tetrac+T4 group[(0.060 ± 0.016) vs.(0.243 ± 0.006),P＜0.05].Conclusion Through binding to membrane integrin αvβ3,thyroid hormone promotes the proliferation of tumor cells by activating the ERK1/2 signaling pathway,and promotes angiogenesis by activating the PKC/PKD1/HDAC5 signaling pathway.