Gu P, Liu H L, Deng Y X, Gao W, Liu J J, Huang X F, Li X Y, Huang H Y
School of Public Health, Guangxi Medical University, Nanning 530021, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2017 Mar 6;51(3):257-261. doi: 10.3760/cma.j.issn.0253-9624.2017.03.013.
To explore the expression of epidermal growth factor receptor(EGFR) protein during benzo(a)pyrene (BaP) induced carcinogenesis. This study, we firstly utilized immunofluorescence assay and Western-blot to examine EGFR expression of the BaP which was constructed previously by project team induced malignant transformation human bronchial epithelial cell (BTC) and the control (16HBE cell). Then, we selected 36 healthy SD rats, divided into two groups according to simple random method, 18 rats each group. The constructed rat lung neoplasm model induced by pulmonary injection of BaP (10 mg/ml of BaP solution in 0.2 ml corn oil), contrast group use 0.2 ml corn oil, lung tissue was collected and the EGFR expression of lung tissue was detected by immunofluorescence assay and Western blot. T analysis was used to test the different of EGFR between two groups. Immunofluorescence analysis showed that the EGFR expression in BTC was significantly higher than 16HBE cell. Meanwhile, Western blot also was used to confirmed this result, the relative expression of EGFR protein in the rats of the model group the control group were 1.04±0.13 and 2.32±0.12, respectively, and the difference was statistically significance (12.39, 0.001). In vivo, well-defined tumor was found in the rat with pulmonary injection of BaP, and the lung showed diffuse alveolar septal thickening, alveolar wall destruction and pulmonary alveoli fusion, which suggested that the rat lung neoplasm model was constructive successfully. Furthermore, we found the EGFR expression of lung was increased dramatically in the rat lung neoplasm model by immunofluorescence detection and Western blot. The relative expression of EGFR protein in the rats of the model group the control group were 0.21±0.03 and 1.30±0.07, respectively, and the difference was statistically significance (12.84, 0.001). Expression of EGFR protein was increased during BaP carcinogenesis, and EGFR may play an important role in the carcinogenesis of BaP.
为探讨表皮生长因子受体（EGFR）蛋白在苯并（a）芘（BaP）诱导致癌过程中的表达情况。在本研究中，我们首先利用免疫荧光分析和蛋白质免疫印迹法检测项目团队先前构建的经BaP诱导恶性转化的人支气管上皮细胞（BTC）及对照组（16HBE细胞）中EGFR的表达。然后，选取36只健康的SD大鼠，采用简单随机法分为两组，每组18只。通过肺内注射BaP（10mg/ml的BaP溶液溶于0.2ml玉米油中）构建大鼠肺癌模型，对照组注射0.2ml玉米油，收集肺组织，采用免疫荧光分析和蛋白质免疫印迹法检测肺组织中EGFR的表达。采用t检验分析两组间EGFR的差异。免疫荧光分析显示，BTC中EGFR的表达显著高于16HBE细胞。同时，蛋白质免疫印迹法也证实了这一结果，模型组大鼠与对照组大鼠EGFR蛋白的相对表达量分别为1.04±0.13和2.32±0.12，差异具有统计学意义（t = 12.39，P = 0.001）。在体内，肺内注射BaP的大鼠出现明确的肿瘤，肺表现为弥漫性肺泡间隔增厚、肺泡壁破坏及肺泡融合，提示大鼠肺癌模型构建成功。此外，通过免疫荧光检测和蛋白质免疫印迹法发现，大鼠肺癌模型中肺组织EGFR的表达显著增加。模型组大鼠与对照组大鼠EGFR蛋白的相对表达量分别为0.21±0.03和1.30±0.07，差异具有统计学意义（t = 12.84，P = 0.001）。EGFR蛋白表达在BaP致癌过程中增加，EGFR可能在BaP致癌过程中起重要作用。
