维甲酸对甲状腺癌ASC蛋白的影响  

The effect of retinoie acid on expression of ASC protein in thyroid cancer

作　　者：薛晓婕[1] 

机构地区：[1]湖北省黄石市中心医院医学检验科,4350001

出　　处：《中华内分泌外科杂志》2012年第1期11-14,共4页Chinese Journal of Endocrine Surgery

基　　金：湖北省自然科学基金项目（2010CDZ054）

摘　　要：目的探索维甲酸诱导甲状腺癌细胞的凋亡情况及凋亡相关斑点样蛋白（ASC）的表达。方法甲状腺滤泡状癌细胞株FIE-133（A组），乳头状甲状腺癌细胞株W3（B组），未分化甲状腺癌细胞株8505C（c组），3组细胞用维甲酸刺激24h共同培养。流式细胞术观察细胞活性，RT—PCR和Western blotting检测ASCmRNA及蛋白的表达。采用双向电泳分离蛋白质，采用PDQuest2-DE软件分析维甲酸＋A组与A组细胞2组间差异表达的蛋白质斑点，并用Western blotting进一步验证。结果流式细胞术结果，维甲酸均导致3组细胞凋亡，A组的凋亡率强于B、C2组（P〈0．01）。RT—PCR和Western blotting检测的ASC mRNA及蛋白的表达，A、B、C3组均有表达，A组强于B、C2组（P〈0．05）。维甲酸作用A组，质谱鉴定了ASC蛋白．Western blotting验证蛋白质ASC与双向电泳的结果一致。结论为维甲酸治疗甲状腺癌的蛋白质研究提供实验资料。Objective To testify the effects of retinoic acid on controlling the proliferation and differentiation of thyroid arcinoma cells and to investigate the role of ASC （ apoptosis-associated specklike protein containing a CARD） in thyroid neoplasm induced by retinoic acid. Methods FIE-133 （group A）, W3 （group B） and 8505C （group C ） were suspended in RPMI 1640 and cultured in the presence of retinoic acid for 24 hours. The activity of thyroid areinoma cells induced by retinoic acid was examined by flow eytometric analysis. The expression level of ASC mRNA and protein was examined by RT-PCR and Western blotting. FIE-133 proteins were separated by 2-DE （two-dimensional electrophoresis） and then stained by silver staining. PDQuest software was used to analyze differentially expressed proteins between group A and group A induced by retinoic acid. Western blotting was used to further identify the protein. Results Flow cytometric analysis showed cells in the 3 groups were induced apoptosis by retinoic acid. The apoptosis ratio was higher in group A than in group B and C（P 〈 0. 01 ）. ASC mRNA and protein were expressed in all the 3 groups and they were higherly expressed in group A than in group B and C （P 〈 0.05）. In group A induced by retinoic acid, ASC protein was ascertained by PD Quest software and the result of Western blotting and 2-DE was accordant. Conclusion Our result provides data for studying proteins in thyroid neoplasm induced by retinoic acid.

关 键 词：乳头状甲状腺癌细胞 未分化甲状腺癌细胞 甲状腺滤泡状癌细胞 凋亡相关斑点样蛋白 蛋白印迹法 流式细胞术 

分 类 号：R736.1[医药卫生—肿瘤]