蛋白激酶CK2抑制剂TBB对甲状腺鳞癌细胞株SW579 CK2β和Akt表达的影响

J4 ›› 2012, Vol. 38 ›› Issue (2): 276-280.

蛋白激酶CK2抑制剂TBB对甲状腺鳞癌细胞株SW579 CK2β和Akt表达的影响

刘超|刘洋|赵颂|王翠瑶|肖建英

辽宁医学院生物化学与分子生物学教研室,辽宁锦州 121001

收稿日期:2011-11-01 出版日期:2012-03-28 发布日期:2012-03-28
通讯作者: 肖建英(Tel:0416-4673239，E-mail:lyxiaojy@163.com) E-mail:lyxiaojy@163.com
作者简介:刘超(1978-)|女|辽宁省铁岭市人|讲师|医学硕士|主要从事肿瘤分子生物学研究。
基金资助:
辽宁省教育厅重点实验室项目资助课题(LS2010101)

Effects of TBB on expressions of CK2&beta|and Akt in thyroid squamous cell carcinoma cell line SW579

LIU Chao,LIU Yang,ZHAO Song,WANG Cui-yao,XIAO Jian-ying

Department of Biochemical and Molecular Biology,Liaoning Medial University,Jinzhou 121001,China

Received:2011-11-01 Online:2012-03-28 Published:2012-03-28

摘要/Abstract

摘要：

目的：观察四溴苯三唑(TBB)对甲状腺鳞癌SW579细胞株CK2β和Akt表达的影响，探讨TBB抗甲状腺癌的作用机制。方法：体外培养SW579细胞，实验分为对照组（未加任何处理组和DMSO组）、TBB组（终浓度为25 μmol/L）和Wortmannin组(终浓度为100 nmol/L)。MTT比色法测定细胞生长抑制率，同时测定蛋白激酶CK2的活性。采用RT-PCR方法检测SW579细胞CK2β及Akt的mRNA表达，分析各处理组CK2β及Akt基因表达变化。采用Western blotting方法检测SW579细胞CK2β及Akt的蛋白表达，分析各处理组CK2β及Akt的蛋白表达变化。结果：与对照组比较，TBB组SW579细胞生长抑制率明显增加（P<0.01），细胞内CK2的活性明显降低（P<0.01）。RT-PCR结果表明，与对照组比较，TBB组CK2β的mRNA 表达水平显著降低（P<0.01），Akt的mRNA 表达水平无明显变化（P>0.05）。Western blotting结果表明，与未处理组比较，Wortmannin组内源性CK2的活性无明显变化（P>0.05），TBB组CK2β的表达明显降低（P<0.01）,同时Akt Thr308和Ser473磷酸化状态受到抑制（P<0.01），总Akt水平无明显变化。结论：TBB可降低SW579细胞蛋白激酶CK2β的表达，进而再下调Akt的活性。

关键词: 四溴苯三唑；甲状腺鳞癌；CK2β；Akt/PKB；SW579

Abstract:

Objective To observe the effects of 4,5,6,7-tetrabromobenzotriazole(TBB) on the expressions of CK2β and Akt in thyroid squamous cell carcinoma cell line (SW579) and to explore their mechanisms.Methods SW579 cells were cultured in vitro and divided into con
trol group,DMSO group，TBB (25 μmol/L) group， and Wortmannin (100 nmol/l group randomly.The inhibitory rate of the growth of SW579 cells was detected with MTT and the protein kinase activity was measured.The CK2β and Akt mRNA and protein expression levels were determined by RT-PCR and Western blotting.Results Compared with control group,the inhibitory rate of growth of SW579 in TBB group was increased（P<0.01） and the cellular activity of CK2 was decreased（P<0.01）. Compared with control group,the expression levels of CK2β mRNA and protein in TBB group were significantly reduced（P<0.01）,while Akt mRNA and protein levels were unchanged（P>0.05）,but phosphorylation status of Thr308 and Ser473 of Akt in TBB group was inhibited（P<0.01）.Conclusion TBB can inhibit the expression and activity of CK2β and down-regulate the activity of Akt of SW579 cells.

Key words: 4,5,6,7-tetrabromobenzotriazole；thyroid squamous cell carcinoma；CK2β；Akt/PKB； SW579

中图分类号:

R581.3

刘超|刘洋|赵颂|王翠瑶|肖建英 . 蛋白激酶CK2抑制剂TBB对甲状腺鳞癌细胞株SW579 CK2β和Akt表达的影响[J]. J4, 2012, 38(2): 276-280.

LIU Chao,LIU Yang,ZHAO Song,WANG Cui-yao,XIAO Jian-ying . Effects of TBB on expressions of CK2&beta|and Akt in thyroid squamous cell carcinoma cell line SW579[J]. J4, 2012, 38(2): 276-280.