Song Li, Wang Yue, Wang Jun, Yang Fan, Li Xiaojun, Wu Yonghui
Department of Occupational and Environmental Health, School of Public Health, Harbin Medical University, Harbin, Heilongjiang, China (mainland).
Department of Occupational Poisoning, Heilongjiang Institute of Labor, Health, and Occupational Disease, Harbin, Heilongjiang, China (mainland).
Med Sci Monit. 2015 Nov 9;21:3434-41. doi: 10.12659/msm.894169.
This study aims to describe trinitrotoluene (TNT)-induced endoplasmic reticulum stress (ERS) and apoptosis in HePG2 cells.
HePG2 cells were cultured in vitro with 0, 6, 12, or 24 μg/ml TNT solution for 12, 24, and 48 h. Western blotting was performed to detect intracellular ERS-related proteins, including glucose-regulated protein (GRP) 78, GRP94, Caspase 4, p-Jun N-terminal kinase (JNK), and C/EBP homologous protein (CHOP). Real-time PCR was used to measure mRNA expression from the respective genes.
The expressions of ERS-related proteins GRP78 and GRP94 as well as mRNA and protein expression of ERS signaling apoptotic CHOP in the TNT treatment group were significantly increased. In addition, the mRNA and protein expression levels of ERS-induced apoptotic protein Caspase-4 were significantly increased. Flow cytometry revealed that after TNT treatment, the apoptosis rate also significantly increased.
TNT could increase the expression levels of GRP78, GRP94, Caspase-4, and CHOP in HePG2 cells; this increase in protein expression might be involved in HePG2 apoptosis through the induction of the ERS pathway.
本研究旨在描述三硝基甲苯（TNT）诱导的人肝癌细胞系（HePG2）内质网应激（ERS）和细胞凋亡情况。
将HePG2细胞分别用0、6、12或24μg/ml的TNT溶液在体外培养12、24和48小时。采用蛋白质免疫印迹法检测细胞内与ERS相关的蛋白，包括葡萄糖调节蛋白（GRP）78、GRP94、半胱天冬酶4（Caspase 4）、磷酸化c-Jun氨基末端激酶（p-Jun N-terminal kinase，p-JNK）和C/EBP同源蛋白（C/EBP homologous protein，CHOP）。使用实时荧光定量PCR检测各基因的mRNA表达。
TNT处理组中ERS相关蛋白GRP78和GRP94的表达以及ERS信号凋亡相关蛋白CHOP的mRNA和蛋白表达均显著增加。此外，ERS诱导的凋亡蛋白Caspase-4的mRNA和蛋白表达水平也显著增加。流式细胞术显示，TNT处理后，细胞凋亡率也显著增加。
TNT可增加HePG2细胞中GRP78、GRP94、Caspase-4和CHOP的表达水平；这种蛋白表达的增加可能通过诱导ERS途径参与HePG2细胞凋亡。
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