摘要
目的观察ERK1/2 MAPK通路在食管鳞状细胞癌(ESCC)Eca109细胞系增殖和迁移中的作用并探讨其作用机制。方法使用MAPK(ERK1/2)抑制剂U0126处理Eca109细胞;细胞计数检测细胞增殖;倒置显微镜下观察细胞形态;实时荧光定量PCR(qRT-PCR)检测ERK1/2 mRNA;Western blot检测总ERK1/2(t-ERK1/2)和磷酸化ERK1/2(p-ERK1/2);MTT和细胞划痕实验检测细胞增殖和迁移能力;qRT-PCR检测MicroRNA-21(miR-21)。结果20μmol/L浓度的U0126可抑制Eca109细胞生长(P<0.05),破坏细胞正常形态,ERK1/2 MAPK通路的活化在蛋白质水平被抑制(P<0.05),Eca109细胞增殖和迁移明显减弱(P<0.05),显著下调miR-21的表达(P<0.05)。结论ERK1/2 MAPK信号通路抑制可减弱Eca109细胞增殖和迁移,其可能的作用机制之一与其抑制miR-21表达有关。
Objective To investigate and migration of Ecal09 cell line in the role and mechanism of ERK1/2 MPAK pathway in regulating proliferation vitro. Methods We treated Ecal09 cells with MAPK（ERK1/2）inhibitor （U0126） ; cell counting method detected the cell proliferation; Cell morphology was observed under inverted micro scope ;qRT-PCR detected ERK1/2 mRNA level; Western blot determined the expression of ERK1/2 protein level; MTI＇ and Wound scratch assay detected cell proliferation and migration; qRT-PCR examined miR-21 expression. Results Cell growth was inhibited by U0126 at 20 p, mol/L doses（P 〈0. 05） , destroyed the normal cell morpholo gy; inaetivited ERK1/2 MPAK pathway at protein level（P 〈 0. 05）; inhibition of ERK1/2 MPAK pathway re- pressed cell proliferation and migration of Ecal09 cells and downregulated the expression of miR-21 （ P 〈 0.05 ）. Conclusions Inhibition of ERK1/2 MPAK pathway inhibits proliferation and migration of Ecal09 cells, one of the most possible mechanism is associated it＇s effect on miR-21 expression.
出处
《基础医学与临床》
CSCD
北大核心
2014年第3期345-349,共5页
Basic and Clinical Medicine
基金
新疆自治区科技厅面上项目(2012211A080)
