PRDX1抑制蛋白酶体抑制剂诱导人甲状腺癌细胞凋亡机制探讨  

Molecular mechanism of PRDX1 attenuates apoptosis induced by proteasome inhibitor in thyrioid carcinoma cell lines

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

机构地区：[1]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 [2]中国医科大学附属第一医院老年病科,辽宁沈阳110001 [3]中国医科大学附属第一医院内分泌科,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2014年第12期899-902,908,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：辽宁省博士科研启动项目(20111110)

摘　　要：目的:初步探讨过氧化还原蛋白1(peroxirodoxin 1,PRDX1)抑制蛋白酶体抑制剂诱导人甲状腺癌细胞凋亡的机制。方法:选取人甲状腺癌细胞,设立随机序列核酸siRNA、siASK1、siPRDX1和siASK1+siPRDX1组,分别用培养液、lactacystin和MG132培养细胞;用蛋白印迹法检测PRDX1、ASK1、p38、JNK1/2、p-ASK1、p-P38和p-JNK1/2在各组甲状腺癌细胞中的表达;用微小RNA干扰技术转染细胞;应用流式细胞仪检测细胞凋亡率。结果:MG132处理组中,p-ASK1是对照组的1.5倍(F=214.14,P<0.001),其下游效应子p-p38和p-JNK1/2分别为对照组的1.34(F=216.75,P<0.001)和1.94倍(F=1 601.68,P<0.001);siASK1下调MG132介导的p-ASK1为随机序列组的0.58倍(F=1 136.82,P<0.001),p-p38为随机序列组的0.73倍(F=507.00,P<0.001),p-JNK1/2为随机序列组的0.51倍(F=6 087.48,P<0.001)。甲状腺癌细胞凋亡率降低为34.25%,显著低于于随机序列组的51.98%,F=18.39,P=0.013。MG132处理组中,siPRDX1组甲状腺癌细胞凋亡率为68.99%显著高于随机序列组的49.58%和siPRDX1+siASK1联合组的41.28%,而siASK1组的甲状腺癌细胞凋亡率为31.85%,显著低于上述两组,组间差异有统计学意义,F=30.13,P<0.001。结论:PRDX1通过负向调节ASK1活性及ASK1-p38和ASK1-JNK通路,抑制ASK1介导的细胞凋亡进而消减MG132对甲状腺癌细胞毒性效应。OBJECTIVE：To investigate molecular mechanism of peroxiredoxins （PRDX1） in inhibiting apoptosis of thyroid cancer cells affected by proteasome inhibitor. METHODS： A panel of cancer cells were selected for the investigation, set up scramble siRNA, siASK1, siPRDX1, siASK1 and siPRDX1, followed by treatment with vehicle control, MG132 or lactacystin; the expression of PRDX1, ASK1, P38, JNK1/2, p-ASK1, p-P38, p-JNK1/2 protein in each group of thyroid cancer cells was confirmed by Western blot,respectively. The small interfering RNA （siRNA） was transfected to thyroid cancer cells;flow cytometry was used to measure apoptotic cells. RESULTS：The protein expressions of p-ASKl,p-38 and p-JNK1/2 were significantly increased in MG132,1.5,1.34 and 1.94 times higher in MG132 treated group compared to those in control group（F = 214. 14, F = 216. 75, F = 1 601. 68, all P〈 0. 001 ）. Compared with the groups treated with scramble,the expressions of p ASK1 in the MG132 treated was also inhibited by 0.58 times（F= I 136.82, P〈0. 001）. The expressions of p-p38 and p-JNK1/2 were also suppressed by 0.73 and 0.51 times in the MG132 treated（F=507.00, F=6 087.48,both P〈0. 001）. The apoptosis rate of thyroid cancer ceils in the MG132 treated was 34.25% which was Significantly lower than 51.98% （F= 18.39, P〈 0. 013） in scramble groups. Among the MG132 groups, the apoptosis rate of the siPRDX1 treated was 68.99 % ,obviously higher than those of the group treated with scramble （49.58%） and the group treated with siPRDX1 ＋ siASK1 （41.28%）;but for the siASK1 treated, the apoPtotic rate was 31.85%, whichwas significantly lower than those of the two groups （F= 30. 13, P〈0. 001 ）. CONCLUSION： PRDX1 can inhibit apoptosis and attenuat the eytotoxie effect of thyroid cancer cells induced by MG132 via negatively regulating the ativity of ASK1 and the pathways of ASK1- p38 and ASK1-JNK.

关 键 词：蛋白酶体抑制剂 细胞凋亡信号调节激酶1 过氧化还原蛋白1 甲状腺肿瘤 

分 类 号：R736.1[医药卫生—肿瘤]