3种MAPK抑制因子对HepG2细胞中Smad2/3磷酸化的影响  

The effect of three MAPK inhibitors on smad2/3 phosphorylation

作　　者：王玉美[1] 梁佳玉[1] 杨雁[1] 

机构地区：[1]安徽医科大学药理学教研室,合肥230032

出　　处：《安徽医科大学学报》2006年第6期648-650,共3页Acta Universitatis Medicinalis Anhui

基　　金：国家新药基金资助项目(编号:969010567)

摘　　要：目的研究3种MAPK抑制因子对TGF-β1活化的HepG2细胞中Smad2/3磷酸化的影响。方法采用MTT方法检测TGF-β1(9pmol)与HepG2共培养0·5、1、2、4、8、16h后,HepG2细胞增殖,TGF-β1(9pmol,0·5h)诱导HepG2细胞活化,在细胞活化前2h分别加入p38、ERK及JNK抑制因子(PD163169、SP600125、PD98059)与HepG2细胞共培养,Westernblot方法检测3种抑制因子对HepG2细胞内Smad2和Smad3磷酸化的影响。结果TGF-β1(9pmol)作用0·5、1h时无明显促进HepG2细胞增殖作用,但作用2、4、8、16h均明显促进HepG2细胞增殖。TGF-β1(9pmol,0·5h)均可诱导Smad2C、Smad2L、Smad3L磷酸化;JNK抑制因子(3、10μmol)与p38抑制因子(1、3、10μmol)可明显抑制Smad2C、Smad3L磷酸化,ERK抑制因子(1、3、10μmol)对Smad2/3磷酸化无明显影响。结论在HepG2细胞中,TGF-β1可能通过JNK、p38途径促进Smad2/3磷酸化。Objective To study the effect of MAPKs inhibitors on Smad2/3 phosphorylation induced by TGF-β1 in HepG2 cells. Methods The cells were stimuilated by TGF-β1 and treated with three inhibitors specific to JNK, p38 and ERK separately. The HepG2 cells were incubated with TGF-β1 （9 pmol） for 0. 5,1,2,4,8,16 h, then the proliferation of HepG2 cells were analyzed by MTT, and the HepG2 cells were pretreated with JNK inhibitor p38 inhibitor and ERK inhibitor for 2h before the addition of TGF-β1 （9 pmol, 0.5 h）, the effects of inhibitors on Smad2/3 pbosphorylation were analyzed by western blot. Results The proliferation of the cells was not remarkable with TGF-β1 （9 pmol） at 0.5 and 1 h, but significant at 2,4,8,16 h ; the phosphorylation level of Smad2C, Smad2L and Smad3L increased in the cells induced by TGF-β1 （9 pmol） at 0.5 h; the phosphorylation level of Smad2C and Smad3L decreased in the cells treated with JNK inhibitor （3,10 μmol） and p38 inhibitor （ 1,3,10 μmol） ; and ERK inhibitor （ 1,3,10 μmol） had no apparent effect on the pbosphorylation of Smad2/3. Conclusion The phosphorylation of Smad2/3 might be induced by TGFμβ1 via activated JNK and p38 in HepG2 cells.

关 键 词：细胞系 肿瘤 转化生长因子Β 丝裂原激活蛋白激酶激酶类/抑制剂及和拮抗剂 信号传导 

分 类 号：R735.7[医药卫生—肿瘤] R345.57[医药卫生—临床医学]