Objective: To study the effect of particulate matter 2.5 (PM(2.5)) on oncogene expression in human bronchial epithelial (HBE) cells. Methods: HBE cells were selected as the study subjects, and PM(2.5) treatment group (10 μg/ml and 50 μg/ml) , negative control group and positive control group (10 μmol/L Cr(6+)) were set. CCK8 assay was used to test the IC(50) value of PM(2.5). HBE cells were treated with PM(2.5) for 24 h at 10 μg/ml and 50 μg/ml, additionally, cells were treated with blank as negative control, 10 μmol/L Cr(6+) as a positive control for 24 h. After the treatment, mRNA expression of oncogenes including c-myc, c-fos, k-ras and p53 were detected by fluorescent quantitative RT-PCR, the protein expression of oncogenes were detected with western blot. Results: The IC(50) value of PM(2.5) in HBE cells is 70.12 μg/ml. The qRT-PCR data showed that compared with the control group, the expression level of c-myc gene increased by respectively 500.1%、780.7%、305.3% after exposure to 10、50 μg/ml PM(2.5) and positive control group; c-fos gene increased respectively 34.0%、76.7%、131.3% after exposure to 10、50 μg/ml PM(2.5) and positive control group; k-ras gene increased respectively 50.3%、107.0%、49.7% after exposure to 10、50 μg/ml PM(2.5) and positive control group; p53 gene decreased by 28.3%、28.7%、59.7% after exposure to 10、50 μg/ml PM(2.5) and positive control group. The western blot results showed that compared with the control group, c-myc protein increased respectively 29.7%、77.3% after exposure to 50 μg/ml PM(2.5) and positive control group; c-fos protein increased respectively 200.3%、137.0% after exposure to 50 μg/ml PM(2.5) and positive control group; k-ras protein increased respectively 106.3%、130.3%、116.7% after exposure to 10、50 μg/ml PM(2.5) and positive control group; p53 protein decreased by 43.7%、53.3%、52.1% after exposure to 10、50 μg/ml PM(2.5) and positive control group. Conclusion: PM(2.5) could promote the expression of oncogenes in HBE cells, the carcinogenicity of haze might be related to promotion of oncogenes expression induced by PM(2.5).
目的： 探讨大气细颗粒物（PM(2.5)）对人支气管上皮细胞（HBE细胞）癌基因表达水平的影响。 方法： 以HBE细胞为研究对象，设PM(2.5)处理组（10 μg/ml和50 μg/ml）、阴性对照组和阳性对照组（10 μmol/L Cr(6+)）。用CCK8实验检测PM(2.5)半数抑制浓度（IC(50)）；应用荧光定量反转录-聚合酶链反应（qRT-PCR）和蛋白免疫印迹（Western blot）检测c-myc、c-fos、k-ras和p53基因和蛋白表达水平变化。 结果： PM(2.5)的IC(50)为70.12 μg/ml。与阴性对照组比较，10和50 μg/ml PM(2.5)组以及阳性对照组HBE细胞c-myc基因表达水平分别升高500.1%、780.7%、305.3%，c-fos基因表达水平分别升高34.0%、76.7%、131.3%，k-ras基因表达水平分别升高50.3%、107.0%、49.7%，p53基因表达水平分别下降28.3%、28.7%、59.7%，差异均有统计学意义（P<0.05）。与阴性对照组比较，50 μg/ml PM(2.5)组和阳性对照组HBE细胞c-myc蛋白表达水平分别升高29.7%、77.3%，c-fos蛋白表达水平分别升高200.3%、137.0%，差异均有统计学意义（P<0.05）；与阴性对照组比较，10和50 μg/ml PM(2.5)组以及阳性对照组HBE细胞k-ras蛋白表达水平分别升高106.3%、130.3%、116.7%，p53蛋白表达水平分别下降43.7%、53.3%、52.1%，差异均有统计学意义（P<0.05）。 结论： 大气PM(2.5)对HBE细胞中癌基因表达具有明显促进作用，PM(2.5)致癌可能与其促进癌基因表达存在密切关系。.
Keywords: Fine particulate matter (PM(2.5)); Gene expression; Haze; Human bronchial epithelial cells; Oncogenes.