苯并[a]芘作用下HSP70、P53在人胚肺细胞中定位及共表达的研究

Study of the Localization and Coexpression of HSP70 and P53 in HEL Cells Treated by Benzo(a)pyrene

目的:探讨苯并[a]芘(BaP)作用下HSP70、P53在人胚肺细胞(HEL)中的定位及共表达。方法:培养正常人胚肺二倍体细胞,分别以0,10,50,100,200μmol·L-1BaP染毒3h,用双重免疫荧光组织化学法和激光扫描共聚焦显微镜检观察HSP70、P53的表达。结果:HEL细胞经不同浓度的BaP染毒3h后,HSP70在胞浆、胞核均有表达,随BaP染毒浓度的增加,逐渐由细胞浆表达向核转移;而P53表达在胞浆、胞核均呈增加趋势(除200μmol·L-1BaP外),胞核中P53蛋白表达水平略高于胞浆;小于50μmol·L-1BaP处理的HEL细胞,HSP70和P53以细胞浆共表达为主,而BaP浓度大于50μmol·L-1时,主要表现为胞核共表达。结论:BaP染毒的人胚肺细胞HSP70、P53的分布及定位方式具有浓度依赖的趋势,表现为随BaP染毒浓度的增加,HSP70与P53由胞浆共表达转变胞核共表达。

Objective： To investigate the localization and co-expression of heat shock protein 70 （HSP70） and P53 in the human embryonic lung （HEL） cells treated by benzo（a）pyrene （BaP）. Methods： HEL cells were cultured and treated at different concentrations of BaP （0, 10, 50, 100, 200 μmol· L^-1） for 3 hours. The expressions of P53 and HSP70 were detected by immunohisto- chemistry and Confocal Laser Scanning Microscopy technology. Results： The expression of HSP70 was detected in both the nucleus and cytoplasm in BaP-treated HEL cells, and the expression mainly concentrated in the nucleus when the concentration of BaP was increasing. A similar distribution of P53 protein was observed in BaP-stimulated HEL cells, however, the levels of P53 in the nucleus were higher than that in the cytoplasm. The co-expression of HSP70 and P53 was found in the cytoplasm at the dose of less than 50 μmol· L^-1, and in the nucleus at the dose of more than 50 μmol· L^-1BaP. Conclusion： The immunolocalition and distribution of HSP70 and P53 in HEL cells treated by benzo（a）pyrene（BaP） is in a dose-dependent manner, and the localization of co-expression of HSP70 and P53 is changed from the cytoplasm to the nucleus as BaP concentration increasing.