摘要
目的:研究MAPK/ERK信号通路中关键信号分子MEK和ERK在胃癌SGC-7901细胞中的表达及PD98059抑制MAPK/ERK通路对胃癌细胞生物学功能的影响。方法:体外培养胃癌细胞株SGC-7901,不同浓度(0、25、50、100、200、300和400 mmol/L)PD98059处理24 h后CCK-8法检测细胞增殖率变化;再用0、25、50和100μmol/L PD98059处理24 h后采用实时荧光定量PCR(qPCR)检测MEK和ERK m RNA的表达量;Western blot检测MEK和ERK蛋白的表达;流式细胞术检测细胞周期和凋亡变化。同时设正常胃黏膜上皮GES-1细胞为对照。结果:与正常胃黏膜上皮GES-1细胞相比,胃癌SGC-7901细胞中MEK和ERK mRNA的表达升高,差异具有统计学意义(P<0.05);p-MEK、p-ERK蛋白的表达亦显著升高,差异具有统计学意义(P<0.05)。0～200μmol/L PD98059处理SGC-7901细胞后,细胞增殖率随着抑制剂浓度的升高而降低(P<0.05)。当PD98059浓度处于200～400μmol/L时抑制作用逐渐趋于平稳。0～100μmol/L PD98059作用后MEK、ERK m RNA的表达量低于对照组(P<0.05),随着PD98059浓度升高,ERK mRNA表达量逐渐降低(P<0.05)。Western blot检测结果显示50和100μmol/L PD98059作用后p-MEK1/2、p-ERK1/2蛋白表达降低(P<0.05)。且抑制剂PD98059使胃癌SGC-7901细胞发生G0/G1期阻滞,可诱导细胞凋亡。结论:MAPK/ERK信号通路在胃癌细胞中激活,PD98059通过抑制MAPK/ERK信号通路的活性可影响胃癌细胞的生物学功能。
OBJECTIVE:To investigate expression of key signaling molecules MEK and ERK in the MAPK/ERK signaling pathway and effect of PD98059 on MAPK/ERK pathway’s biological function in the gastric cancer SGC-7901 cells.METHODS:Gastric cancer cell line SGC-7901 was cultured in vitro.Cell proliferation rates were detected by the CCK-8 method after treatment with different concentrations of PD98059 at 0,25,50,100,200,300 and 400μmol/L for 24 h.Expression of MEK and ERK mRNA was detected by real-time quantitative PCR(qPCR)after treatment with 0,25,50 and 100μmol/L PD98059 for 24 h.Expression of MEK and ERK proteins was detected by Western blot.Cell cycle and apoptosis were detected by flow cytometry.The normal gastric mucosal epithelial GES-1 cells were used as controls.RESULTS:Compared with the normal cells,expression of MEK and ERK mRNA and p-MEK and p-ERK proteins in the gastric cancer cells was significantly increased(P<0.05).After treatment with PD98059,the cell proliferation rates were decreased with increasing concentration and in a dose-dependent manner.When the concentration of PD98059 was between 200 and 400μmol/L,the inhibition became stabilized.Expression of the MEK and ERK mRNA was lower than that of the control group after treatment with 0-100μmol/L PD98059(P<0.05).With increasing concentrations of PD98059,expression of ERK mRNA was gradually reduced(P<0.05).Western blot analyses show that expression of p-MEK1/2 and p-ERK1/2 protein decreased after treatment with 50 and 100μmol/L PD98059(P<0.05).Moreover,PD98059 caused G0/G1 phase arrest and induced apoptosis.CONCLUSION:MAPK/ERK signaling pathway was active in the gastric cancer cells.PD98059 inhibited activities in the MAPK/ERK signaling pathway and affected the biological function of the gastric cancer cells.
作者
刘梦琪
张文文
陈晓伟
沈孝兵
LIU Mengqi;ZHANG Wenwen;CHEN Xiaowei;SHEN Xiaobing(Key Laboratory of Environmental Medicine and Engineering of Ministry of Education,School of Public Health,Southeast University,Nanjing 210009,Jiangsu,China)
出处
《癌变．畸变．突变》
CAS
2019年第1期15-21,共7页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金(81172619
81472940)
