PM_(2.5)通过p38MAPK信号通路对HBE细胞癌基因表达的影响

Effects of PM_(2.5) on oncogene expression via p38MAPK signal pathway in HBE cells

目的探讨大气细颗粒物PM_(2.5)通过p38MAPK信号通路对人支气管上皮(human bronchial epithelial,HBE)细胞癌基因表达的影响。方法设立空白对照组、PM_(2.5)组、SB203580组、PM_(2.5)+SB203580组。以10μmol/L的p38MAPK抑制剂SB203580预先处理HBE细胞30 min,接着用无血清DMEM培养基培养24 h作为SB203580组;用50μg/mL PM_(2.5)染毒24 h作为PM_(2.5)组;用10μmol/L的SB203580预先处理HBE细胞30 min,然后用50μg/mL的PM_(2.5)染毒24 h作为PM_(2.5)+SB203580组;应用荧光定量PCR和Western blot检测C⁃MYC、C⁃FOS、K⁃RAS、P53、p38MAPK基因的mRNA和蛋白质表达水平变化。结果与对照组比较,PM_(2.5)组C⁃MYC、C⁃FOS、K⁃RAS、p38MAPK基因表达分别升高54.0%、49.2%、96.1%、74.1%,P53基因表达下降49.1%(均P<0.01)。与PM_(2.5)组比较,PM_(2.5)+SB203580组C⁃MYC、C⁃FOS、K⁃RAS和p38MAPK基因表达分别下降21.4%、20.8%、39.3%和21.4%,P53基因表达升高43.1%(P<0.01或P<0.05)。Western blot结果显示,与对照组相比,PM_(2.5)组C⁃MYC、C⁃FOS、K⁃RAS、p38MAPK蛋白表达分别升高110.6%、46.7%、35.5%、82.8%,P53蛋白表达下降42.7%(均P<0.01)。与PM_(2.5)组比较,PM_(2.5)+SB203580组C⁃MYC、C⁃FOS、K⁃RAS和p38MAPK蛋白表达分别下降37.7%、16.2%、19.6%和25.9%,P53蛋白表达上升43.5%(P<0.01或P<0.05)。结论p38MAPK抑制剂明显影响PM_(2.5)对癌基因表达的作用,提示p38MAPK信号通路在PM_(2.5)致癌基因表达作用中发挥重要作用。

Objective To investigate the effects of PM_(2.5) on oncogene expression via p38MAPK signaling pathway in human bronchial epithelial(HBE)cells.Methods Control group,PM_(2.5) group,SB203580 group and PM_(2.5)+SB203580 group were established.HBE cells were treated with 10μmol/L p38MAPK inhibitor SB203580 for 30 min,then cultured in serum‐free DMEM medium for 24 h as SB203580 group;HBE cells were treated with 50μg/mL PM_(2.5) for 24 h as PM_(2.5) group;HBE cells were treated with10μmol/L SB203580 for 30 min,then exposed to 50μg/mL of PM_(2.5) for 24 h as the PM_(2.5)+SB203580 group.The mRNA and protein expression levels of C⁃MYC,C⁃FOS,K⁃RAS,P53 and p38MAPK were detected by real‐time PCR and Western blot.Results PCR results showed that compared with the control group,the mRNA expression of C⁃MYC,C⁃FOS,K⁃RAS,p38MAPK in the PM_(2.5) group increased by 54.0%,49.2%,96.1%,74.1%,and P53 decreased by 49.1%.Compared with the PM_(2.5) group,the mRNA expression of C⁃MYC,C⁃FOS,K⁃RAS and p38MAPK in the PM_(2.5)+SB203580 group decreased by 21.4%,20.8%,39.3%,21.4%,and P53 increased by 43.1%(P<0.01 or P<0.05).Western blot results showed that compared with the control group,the protein expression levels of C⁃MYC,C⁃FOS,K⁃RAS,p38MAPK in the PM_(2.5) group increased by 110.6%,46.7%,35.5%,82.8%,and P53 decreased by 42.7%(all P<0.01).Compared with the PM_(2.5) group,the protein expression levels of C⁃MYC,C⁃FOS,K⁃RAS,p38MAPK in the PM_(2.5)+SB203580 group decreased by 37.7%,16.2%,19.6%,25.9%,and P53 increased by 43.5%(P<0.01 or P<0.05).Conclusion The p38MAPK inhibitor obviously influenced the effects of PM_(2.5) on oncogene ex‐pression,indicating that p38MAPK signaling pathway plays an important role in PM_(2.5)-induced oncogene expression.