ERK1/2介导的bFGF在乳腺癌细胞系MCF-7增殖中的作用

Effects of bFGF on proliferation in breast carcinoma MCF-7 cell line via ERK1/2 pathway

目的观察Ras-Raf-ERK1/2途径介导的碱性成纤维细胞生长因子(bFGF)对乳腺癌细胞系MCF-7细胞增殖的影响,探讨bFGF及其信号传导途径与乳腺癌发生及发展的关系。方法应用细胞免疫组织化学技术检测不同浓度bFGF处理细胞后ERK1/2蛋白的表达;应用蛋白印迹技术检测不同浓度、不同时间bFGF和bFGF+PD98059处理MCF-7细胞后,细胞外信号调节蛋白激酶(ERK1/2)蛋白表达水平的变化。结果经bFGF不同浓度(0、12.5、25及50ng/mL)处理MCF-7乳腺癌细胞系后,ERK蛋白表达随着浓度的增加而增强,且与bFGF水平呈剂量依赖关系;以最佳工作浓度25ng/mLbFGF处理MCF-7乳腺癌细胞系不同时间后,其表达活性明显高于对照组和bFGF+PD98059组。加入bFGF抑制剂PD98059后,bFGF诱导MCF-7细胞系p-ERK1/2活性的作用受到抑制,ERK蛋白表达明显下调。结论 bFGF通过Ras-Raf-ERK1/2途径的介导,促进了MCF-7细胞的增殖,进而抵抗无血清诱导的凋亡。抑制剂PD98059可阻断此诱导作用,使ERK表达明显下调。ERK信号传导通路可能是乳腺癌侵袭和转移的重要途径,ERK1/2和bFGF的表达可为乳腺癌治疗的靶点提供理论依据。

[Objective]To investigate the effect of basic fibroblast growth factor （bFGF）on the proliferation in breast Carcinoma cell line MCF-7 via ERK1/2 signal transduction pathway. To study the correlation of basic fibroblast growth factor （bFGF） to Extra cellular signal-regulated kinase （ERK） expressions in breast carcinoma and cell line MCF-7.[Methods]Cell immunohistochemical method was used to detect the expression of ERK in MCF-7 cell via treated bFGF. Western blot was used to detect the protein levels of ERK1/2 in breast carcinoma MCF-7 cells that was treated with bFGF alone （bFGF group） and bFGF and PD98059 （PD98059＋bFGF group）, respectively. [Result]The bFGF different concentrations processing MCF-7 breast cancer lines, ERK protein expression increased with the increase of bFGF concentration （0, 12.5, 25, 50 ng/mL） and in a dose-dependent way. The expression of ERK activity was obviously higher than the control group and PD98059＋ bFGF group, when MCF-7 breast cancer lines were treated with the best work concentration of FGF 25 ng/mL. After joining bFGF inhibitors PD98059, the role of bFGF induction of p-ERK1/2 activity was restrained, and ERK protein expression. was obviously down-regulated. [Conclusions] ERK1/2 signal transduction pathway might improve breast cancer progression by bFGF expression, and might play an important roles in invasion and metastasis of breast cancer.