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25μmol·L-1)和Wortmannin组(终浓度为100 nmol·L-1)。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。
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 SW579cells were cultured in vitro and divided into control group, DMSO group, TBB （25 μmol · L ^-1 ） group, and Wortmannin （100 nmol · L^-1） group randomly. The inhibitory rate of the growth of SW579 cells was detected with MTT and the protein kinase activity was measured. The CK213 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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2012年第2期276-280,共5页
Journal of Jilin University：Medicine Edition
基金
辽宁省教育厅重点实验室项目资助课题(LS2010101)
