激活蛋白激酶A信号通路对甲状腺鳞癌SW579细胞株的生长抑制作用  

Effects of proliferation in thyoid squamous cell carinoma SW579 by activating the signal pathway of PKA

作　　者：高月[1] 肖建英[1] 赵颂[1] 张秀梅[1] 刘超[1] 王翠瑶[1] 贾昂[1] 

机构地区：[1]辽宁医学院生物化学及分子生物学教研室,辽宁锦州121001

出　　处：《中国现代医学杂志》2012年第17期31-34,共4页China Journal of Modern Medicine

基　　金：辽宁省教育厅重点实验室项目(No:LS2010101)

摘　　要：目的探讨蛋白激酶A(protein kinase A,PKA)信号通路对甲状腺鳞癌SW579细胞株的生长抑制作用及机制。方法利用PKA特异性激活剂双丁酰环化腺苷酸(dbcAMP)0.5、1.0和2.0 mmol/L作用于甲状腺鳞癌SW579细胞24、48和72 h后用四甲基偶氮唑蓝比色法(MTT法)检测SW579细胞的生长活性;酶联免疫吸附法(ELISA法)检测经dbcAMP 1.0 mmol/L作用48 h后PKA、成熟促进因子(message processing fac-tor,MPF)的活性;免疫印迹法(Western blotting法)检测经dbcAMP 1.0 mmol/L作用48 h CDC25B-pS323(celldivision cycle 25B-pS323)磷酸化水平。结果 MTT法结果显示,在dbcAMP作用下,SW579细胞活力呈逐渐下降趋势,随dbcAMP剂量的增加和作用时间的延长,其作用越明显。ELISA法显示,dbcAMP使PKA活性升高而使MPF活性下降。未加dbcAMP组PKA、MPF活性分别为(32.5±2.49)nmol/L和(115.5±7.53)ng/L;dbcAMP组PKA、MPF活性分别为(436.33±12.85)nmol/L和(22.87±2.57)ng/L,两组间比较差异有统计学意义(P<0.01)。Western blotting显示,dbcAMP能够使CDC25B-pS323磷酸化水平上调。未加dbcAMP组和dbcAMP组比较差异有统计学意义(P<0.01)。结论利用dbcAMP激活PKA信号通路,可以抑制甲状腺鳞癌SW579细胞生长增殖,其抑制作用可能与PKA/CDC25B/MPF信号通路的调控有关。【Objective】 To study the effect of PKA signal pathway on growth inhabition.【Methods】 SW579 cells were treated with the specific PKA agent activation dbcAMP（0.5,1.0,2.0 mmol/L） at 24,48,72 h.The growth activity of cells was detected by MTT;the activity of PKA and MPF through the ELISA;the phosphorylation level of CDC25B-S323 by Western Blot.【Results】 MTT showed that the growth activity of SW579 cells was reduced;ELISA showed that dbcAMP could increase the activity of PKA and decrease the activity of MPF.The activities of PKA and MPF were respectively（32.5±2.49） nmol/L and（115.5±7.53） ng/L in group without dbcAMP while the activities of PKA and MPF were respectively（436.33±12.85） nmol/L and（22.87±2.57） ng/L in group with dbcAMP.The difference was significant between the two groups.Western blot showed that CDC25B-pS323 increased.Cdc25B-pSer323/β-actin was（0.050±0.006） in group without dbcAMP while Cdc25B-pSer323/β-actin was（0.148±0.014） in group with dbcAMP.The difference was significant between the two groups.【Conclusions】 dbcAMP can inhibit the proliferation of SW579 cells.The mechanism on inhibiting the growth of thyroid cancer maybe related to the PKA/CDC25B/MPF signal path.

关 键 词：甲状腺鳞癌 蛋白激酶A 成熟促进因子 

分 类 号：R736.1[医药卫生—肿瘤]