PD98059抑制MAPK/ERK信号通路对胃癌细胞生物学功能的影响

doi:10.3969/j.issn.1004-616x.2019.01.003

癌变·畸变·突变 ›› 2019, Vol. 31 ›› Issue (1): 15-21.doi: 10.3969/j.issn.1004-616x.2019.01.003

PD98059抑制MAPK/ERK信号通路对胃癌细胞生物学功能的影响

刘梦琪, 张文文, 陈晓伟, 沈孝兵

东南大学公共卫生学院环境医学工程教育部重点实验室, 江苏南京 210009

收稿日期:2018-08-08 修回日期:2018-10-17 出版日期:2019-01-31 发布日期:2019-01-31
通讯作者: 沈孝兵,E-mail:xb.shen@seu.edu.cn E-mail:xb.shen@seu.edu.cn
作者简介:刘梦琪,E-mail:1399821218@qq.com。
基金资助:
国家自然科学基金（81172619，81472940）

Inhibitory effects of PD98059 on the MAPK/ERK signaling pathway in gastric cancer cells

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

Received:2018-08-08 Revised:2018-10-17 Online:2019-01-31 Published:2019-01-31

摘要/Abstract

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100 μmol/L PD98059处理24 h后采用实时荧光定量PCR（qPCR）检测MEK和ERK mRNA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）。0~200 μmol/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200~400 μmol/L时抑制作用逐渐趋于平稳。0~100 μmol/L PD98059作用后MEK、ERK mRNA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100 μmol/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。

关键词: 胃癌, MAPK/ERK信号通路, PD98059, 细胞增殖, 细胞凋亡

Abstract: 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 400 μmol/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 100 μmol/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 400 μmol/L,the inhibition became stabilized. Expression of the MEK and ERK mRNA was lower than that of the control group after treatment with 0-100 μmol/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 100 μmol/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.

Key words: gastric cancer, MAPK/ERK signaling pathway, PD98059, cell proliferation, apoptosis

中图分类号:

R730.2

刘梦琪, 张文文, 陈晓伟, 沈孝兵. PD98059抑制MAPK/ERK信号通路对胃癌细胞生物学功能的影响[J]. 癌变·畸变·突变, 2019, 31(1): 15-21.

LIU Mengqi, ZHANG Wenwen, CHEN Xiaowei, SHEN Xiaobing. Inhibitory effects of PD98059 on the MAPK/ERK signaling pathway in gastric cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 15-21.

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PD98059抑制MAPK/ERK信号通路对胃癌细胞生物学功能的影响

Inhibitory effects of PD98059 on the MAPK/ERK signaling pathway in gastric cancer cells

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