人胰腺癌细胞株HPAC和BxPC-3中Src/MAPK信号转导途径中pp60c-Src和pERK1/2表达的差异

目的研究人胰腺癌细胞株HPAC和BxPC-3中Src/MAPK信号转导系统,探寻不同类型胰腺癌信号转导通路的差异,为研究靶点药物提供基础。方法将1640培养的HPAC和BxPC-3经细胞计数相同后各分为4组,分别为对照组、TGF-α组、TGF-α+PP2组和TGF-α+PD组。细胞贴壁并达70%融合后,换无血清培养48h。无血清处理作对照,用无血清培养基溶解终浓度为7nmol/L的TGF-α刺激TGF-α组2h,TGF-α+PP2组先加入终浓度为10μmol/L的PP2刺激细胞20min后加入7nmol/L的TGF-α刺激2h,TGF-α+PD组先加入30μmol/L的PD98059刺激细胞20min后加入终浓度为7nmol/L的TGF-α刺激2h。用蛋白印迹方法分析HPAC和BxPC-3的4组细胞pp60c-Src和pERK1/2的表达。结果HPAC细胞pp60c-Src的表达:对照组显著高于TGF-α组、TGF-α+PD组(P<0.05)和TGF-α+PP2组(P<0.01);HPAC细胞pERK1/2的表达:TGF-α组显著高于对照组和TGF-α+PD组(P<0.01),与TGF-α+PP2组相比较差异无统计学意义(P>0.05)。BxPC-3细胞pp60c-Src的表达:TGF-α组显著高于对照组(P<0.05)和TGF-α+PP2组(P<0.01),与TGF-α+PD组相比较差异无统计学意义(P>0.05);BxPC-3细胞pERK1/2的表达:TGF-α组显著高于对照组、TGF-α+PD组和TGF-α+PP2组(P<0.01)。结论HPAC和BxPC-3细胞中Src/MAPK信号途径存在差异。推测在BxPC-3细胞株中c-Src是活化Ras/Raf/MAPK途径的关键信号分子,应进一步研究在HPAC细胞株中c-Src的作用。

Differential Expressions of pp60c-Src and pERK1/2 in Src/MAPK Signal Transduction Pathway of Human Pancreatic Cancer Cell Line HPAC and BxPC-3

Objective To acquire anti tumor target drug basic research data,we investigated the difference in Src/MAPK signal transduction pathway of human pancreatic cancer cell line HPACand BxPC-3.Methods HPACand BxPC-3 were cultured in RPMI 1640 and the same cell number divided into four groups:control,TGF-α,TGF-α+PP2 and TGF-α+PD.Cells were changed to serum free medium in condition of cell adhesion and 70% confluency and cultured 48 h.Control were treated by serum free medium;TGF-α group were treated 2 h by final concretion 7nmol/L TGF-α in serum free medium;TGF-α+PP2 group were treated 20 min by final concretion 10 μmol/L PP2 and then 2 h by final concretion 7nmol/L TGF-α;TGF-α+ PDgroup were treated 20 min by final concretion 30 μmol/L PD and then 2 h by final concretion 7 nmol/L TGF-α.The expression of pp60c-Src and pERK1/2 were analyzed by Western bloting.Results The expression of pp60c-Src in cell line HPACin control significantly higher than TGF-α group,TGF-α+PDgroup(P<0.05)and TGF-α+PP2 group(P<0.01);The expression of pERK1/2 in HPACin TGF-α group were significantly higher than that of control and TGF-α+PDgroup(P<0.01),but no significantly difference to that of TGF-α+PP2group(P>0.05).pp60c-Src expression of TGF-α group in BxPC-3 were significantly higher than that of control(P<0.05),and that of TGF-α+ PP2 group(P<0.01),but not significantly different to that of TGF-α+PDgroup(P>0.05);The expression of pERK1/2 of TGF-α group in BxPC-3 were significantly higher than that of control,TGF-α+PDgroup and TGF-α+PP2 group(P<0.01).Conclusion There were some difference in Src/MAPK signal transduction pathway between human pancreatic cancer cell line HPAC and BxPC-3.We supposed c-src is key signal molecular in activation of Ras/Raf/MAPK pathway in human pancreatic cancer cell line BxPC-3.The mechanism of c-src in human pancreatic cancer cell line HPAC should advanced explored.