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目的研究p38丝裂原活化蛋白激酶(MAPK)和细胞外信号调节激酶(ERK)1/2抑制剂对镉染毒NRK肾细胞后原癌基因c-myc与c-fos表达的影响。方法用不同剂量CdCl2(0、2.5、5.0、10.0μmol/L)染毒大鼠NRK细胞12h,p38MAPK抑制剂SB20358010.0μmol/L和ERK1/2抑制剂PD9805910μmol/L预处理NRK细胞0.5h后,再加入10.0μmol/L CdCl2染毒细胞,用流式细胞技术检测细胞凋亡率,SYBRGreenI荧光定量PCR检测c-myc与c-fos mRNA表达。结果流式细胞仪检测NRK细胞染镉后细胞凋亡率随染毒剂量增加而增加;p38MAPK抑制剂SB203580+10.0μmol/L CdCl2组与单独染镉组比较,细胞凋亡率显著下降;但ERK1/2抑制剂PD98058+10.0μmol/LCdCl2组与单独染镉组比较,细胞凋亡率显著升高。实时荧光定量PCR检测NRK细胞染镉后c-myc和c-fosmRNA表达明显增强;单独用SB203580和PD98058刺激NRK细胞后c-myc和c-fos mRNA表达与对照组比较,差异无统计学意义,但SB203580和PD98058预处理NRK细胞0.5h后加入10.0μmol/L CdCl2与单纯10.0μmol/L染镉组比较,c-myc和c-fos mRNA表达显著降低。结论MAPK信号转导通路可能在镉诱导原癌基因c-myc和c-fos表达中发挥作用。
Objective To investigate the effects of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) 1/2 inhibitor on c-myc and c-fos expression in cadmium-exposed NRK kidney cells. Methods NRK cells were exposed to CdCl2 (0,2.5,5.0,10.0μmol / L) for 12h, NRK cells pretreated with p38MAPK inhibitor SB20358010.0μmol / L and ERK1 / 2 inhibitor PD9805910μmol / L for 0.5h, Then 10.0μmol / L CdCl2 was added to the cells, the apoptosis rate was detected by flow cytometry, and the expression of c-myc and c-fos mRNA was detected by SYBR Green I fluorescence quantitative PCR. Results Flow cytometry showed that the apoptotic rate of NRK cells increased with the dose of cadmium, while the apoptosis rate of p38 MAPK inhibitor SB203580 + 10.0 μmol / L CdCl2 group was significantly lower than that of cadmium alone group. However, ERK1 / 2 inhibitor PD98058 + 10.0μmol / LCdCl2 group compared with cadmium alone group, the apoptosis rate was significantly increased. The expression of c-myc and c-fos mRNA in NRK cells was significantly increased by real-time fluorescent quantitative PCR. Compared with the control group, the expression of c-myc and c-fos mRNA in NRK cells stimulated by SB203580 and PD98058 alone had no statistical significance, However, the expressions of c-myc and c-fos mRNA in NRK cells pretreated with SB203580 and PD98058 for 0.5 h were significantly decreased compared with those treated with 10.0 μmol / L CdCl2 and 10.0 μmol / L Cd 2+ alone. Conclusion MAPK signal transduction pathway may play a role in cadmium-induced proto-oncogene c-myc and c-fos expression.