蛋白酶体抑制剂诱导甲状腺癌细胞凋亡机制的初步研究  

Proteasome inhibitor-mediated apoptosis mechanism in undifferentiated thyroid cancer cells

作　　者：都镇先[1] 刘宝琴[2] 张海燕[3] 邓娓娓[3] 王华芹[2] 

机构地区：[1]中国医科大学附属第一医院内分泌科,辽宁沈阳110001 [2]中国医科大学基础医学院生化与分子生物教研室,辽宁沈阳110001 [3]中国医科大学附属第一医院老年病科,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2010年第2期85-88,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30740086);辽宁省科技攻关项目(2007225021)

摘　　要：目的：探讨蛋白酶体抑制剂在诱导甲状腺未分化癌细胞系的凋亡作用中与核转录因子（NFκ-B）活性的关系。方法：利用蛋白酶体抑制剂PSI、EPOX和MG132作用7种甲状腺未分化癌细胞系,MTT法检测细胞生存率、乳酸脱氢酶（LDH）释放检测各组细胞LDH活性,利用酶联免疫吸附（ELISA）法定量分析NF-κBDNA结合活性,Caspases-3酶分析测定活性。结果：ARO和8305C细胞系对蛋白酶体抑制剂完全不敏感,FRO和KTC2细胞系IC50分别为PSI 4-6 nmol/L、EPOX 1-5 nmol/L和MG132 0.5-1μmol/L;KTC1和KTC3细胞系IC50分别为PSI 10-15 nmol/L、EPOX 10-20 nmol/L和MG132 1-2μmol/L;与对照组相比,所有细胞系应用PSI后,NFκ-B DNA结合活性减少,P〈0.05;应用EPOX时,NFκ-BDNA结合活性在FRO、KTC2、KTC3和8505细胞系中减少（P〈0.05）,但在ARO、KTC1和8305C细胞中保持不变（P〉0.05）。应用MG132处理后,NFκ-B DNA结合活性在所有细胞系中均无明显变化,P〉0.05;各组细胞系基础状态下Caspase-3活性差异无统计学意义,P〉0.05,作用后ARO和8305C细胞系Caspase-3活性最低,FRO和KTC2细胞系Caspase-3活性最高,是基础状态的10-12倍。KTC1和KTC3细胞系Caspase-3活性中等,是基础状态的6-7倍。结论：蛋白酶体抑制剂除通过抑制NFκ-B活性以影响甲状腺癌细胞生存之外,尚可能存在其他抗肿瘤细胞生存途径。OBJECTIVE：To investigate the relationship between nuclear factor（NF-κB）activity and proteasome inhibitor-mediated cell death in undifferentiated thyroid cancer cells.METHODS：Seven undifferentiated thyroid cancer cell lines were treated with proteasome inhibitors（PSI,EPOX and MG132）.The cell viability,toxicity and death were measured by MTT assay,LDH release assay and FCM,respectively.DNA binding activity of NF-κB and Caspase-3 activity were analyzed by ELISA.RESULTS：ARO and 8305C cells were completely insensitive to proteasome inhibitors.In FRO and KTC2 cells,the IC50（s） of PSI,EPOX and MG132 were 4-6 nmol/L,1-5 nmol/L and 0.5-1 μmol/L,respectively.In KTC1 and KTC3 cells,the IC50（s） of these reagents were 10-15 nmol/L,10-20 nmol/L and 1-2 μmol/L,resepectively.As compared with the control,PSI reduced DNA binding of NF-κB in all the cell lines（P〈0.05）.EPOX decreased NF-κB activity in FRO,KTC2,KTC3 and 8505C cells（P〈0.05）,whereas had no effect in ARO,KTC1 and 8305C cells（P〈0.05）.MG132 had no obvious effects on NF-κB activity in the panel of cells（P〈0.05）.Under basal condition,Caspase-3 activity had no obvious difference in the panel of cells（P〈0.05）.Upon exposure to proteasome inhibitors,ARO and 8305C cells demonstrated the lowest activity of Caspase-3.FRO and KTC2 cells showed the highest activity of Caspase-3,up to 10-12 folds compared with the control.KTC1 and KTC3 cells showed moderate Caspase-3 activity,being 6-7 folds as compared with the control.CONCLUSION：Except for suppression of NF-κB activity,proteasome inhibitors might modulate thyroid cancer cell viability via affecting additional cell survival/death pathway.

关 键 词：甲状腺肿瘤 蛋白酶体抑制剂 NF-ΚB 细胞凋亡 

分 类 号：R736.1[医药卫生—肿瘤]