GSTM5 nuclear translocation induced by tumor necrosis factor-α_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

ZENG Yanting ^1,2 , LI Li ² , YUAN Weifeng ² , GUO Zhenhui ³ ,  HUANG Wenjie ^1,2

1. Postgraduate Institute of Southern Medical University, Guangzhou, Guangdong 510182, P. R. China;
2. Department of Respiratory Medicine, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou, Guangdong 510010, P. R. China;
3. Department of MICU, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou, Guangdong 510010, P. R. China;

Corresponding author：

HUANG Wenjie, Email: huangyelu1029@vip.163.com

Keywords：

Glutathione S-transferase mu 5; Recombinant lentiviral expression vector; 16HBE; Nuclear translocation; Tumor necrosis factor-α

DOI：

10.7507/1671-6205.201611031

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

ObjectiveTo establish 16HBE cell lines stably expressing glutathione S-transferase mu 5 (GSTM5) gene, and explore the mechanism of GSTM5 nuclear translocation. MethodsRecombinant lentiviral expression vector containing GSTM5 gene was constructed and lentivirus was produced. After lentivirus infection of 16HBE cells, 16HBE-GSTM5 cell lines were obtained by screening with puromycin. Expression of GSTM5 in different cells was examined by RT-qPCR and Western blot. The nuclear translocation of GSTM5 was observed by confocal laser scanning microscope, after the 16HBE-GSTM5 cell lines were treated with tumor necrosis factor-α (TNF-α; 10 ng/ml) for 0.5 hour. ResultsLentiviral expression plasmids, PLVX-puro-3*flag-SBP-GSTM5-C and PLVX-puro-GSTM5-SBP-3*flag-N, were constructed and lentiviral particles were successfully packed. After infected with lentivirus and screened by puromycin, two cell lines, 16HBE-GSTM5-SBP-3*flag-N and 16HBE-3*flag-SBP-GSTM5-C, were obtained. GSTM5 expression in these two cell lines was significantly higher compared with the control group and parental cells. After treated with TNF-α for 0.5 hour, the nuclear translocation of GSTM5 in 16HBE-GSTM5-SBP-3*flag-N was much more obviously than that in 16HBE-3*flag-SBP-GSTM5-C. ConclusionThe N-terminal region of GSTM5 is critical for nuclear translocation induced by TNF-α, which is mediated by a novel and non-classical nuclear localization signal.

Citation： ZENG Yanting, LI Li, YUAN Weifeng, GUO Zhenhui, HUANG Wenjie. GSTM5 nuclear translocation induced by tumor necrosis factor-α. Chinese Journal of Respiratory and Critical Care Medicine, 2017, 16(5): 451-456. doi: 10.7507/1671-6205.201611031 Copy

1.	Prabhakaran P. Acute respiratory distress syndrome. Indian Pediatr, 2010, 47(10): 861-868.
2.	Lee IT, Yang CM. Role of NADPH oxidase/ROS in pro-inflammatory mediators-induced airway and pulmonary diseases. Biochem Pharmacol, 2012, 84(5): 581-590.
3.	Hayes JD, Flanagan JU, Jowsey IR. Glutathione transferases. Annu Rev Pharmacol Toxicol, 2005, 45: 51-88.
4.	赵艳雪, 李理, 袁伟锋, 等. 谷胱甘肽 S 转移酶 mu5 对肿瘤坏死因子 α 诱导人支气管上皮细胞氧化损伤的调节作用研究. 中国呼吸与危重监护杂志, 2016, 15(3): 284-288.0.
5.	ARDS Definition Task Force, Ranieri VM, Rubenfeld GD, Thompson BT, et al. Acute respiratory distress syndrome: the Berlin Definition. JAMA, 2012, 307(23): 2526-2533.
6.	Castillo RL, Carrasco Loza R, Romero-Dapueto C. Pathophysiological approaches of acute respiratory distress syndrome: novel bases for study of lung injury. Open Respir Med J, 2015, 9: 83-91.
7.	邱娴, 胡水旺, 徐俊, 等. A549 细胞炎症氧化应激模型 Nox1 启动子差异结合蛋白的筛选. 南方医科大学学报, 2013, 33(05): 703-707.0.
8.	黄云, 袁伟锋, 李理, 等. 谷胱甘肽硫转移酶 M5 通过 p38MAPK/NF-κB 途径下调肿瘤坏死因子 α 介导的人支气管上皮细胞炎症水平. 中国呼吸与危护杂志, 2016, 15(2): 171-175.0.
9.	Wu X. Transcription start regions in the human genome are favored targets for MLV integration. Science, 2003, 300(5626): 1749-51.
10.	Pandya SE, Klimatcheva V. Lentivirus and foamy virus vectors: novel gene therapy tools. Expert Opin Biol Ther, 2001, 1(1): 17-40.
11.	Kawakatsu M. Nuclear translocation of glutathione S-transferase p is mediatedby a non-classical localization signal. Biochemical and Biophysical Research Communications: p. 745-750.

1. Prabhakaran P. Acute respiratory distress syndrome. Indian Pediatr, 2010, 47(10): 861-868.
2. Lee IT, Yang CM. Role of NADPH oxidase/ROS in pro-inflammatory mediators-induced airway and pulmonary diseases. Biochem Pharmacol, 2012, 84(5): 581-590.
3. Hayes JD, Flanagan JU, Jowsey IR. Glutathione transferases. Annu Rev Pharmacol Toxicol, 2005, 45: 51-88.
4. 赵艳雪, 李理, 袁伟锋, 等. 谷胱甘肽 S 转移酶 mu5 对肿瘤坏死因子 α 诱导人支气管上皮细胞氧化损伤的调节作用研究. 中国呼吸与危重监护杂志, 2016, 15(3): 284-288.0.
5. ARDS Definition Task Force, Ranieri VM, Rubenfeld GD, Thompson BT, et al. Acute respiratory distress syndrome: the Berlin Definition. JAMA, 2012, 307(23): 2526-2533.
6. Castillo RL, Carrasco Loza R, Romero-Dapueto C. Pathophysiological approaches of acute respiratory distress syndrome: novel bases for study of lung injury. Open Respir Med J, 2015, 9: 83-91.
7. 邱娴, 胡水旺, 徐俊, 等. A549 细胞炎症氧化应激模型 Nox1 启动子差异结合蛋白的筛选. 南方医科大学学报, 2013, 33(05): 703-707.0.
8. 黄云, 袁伟锋, 李理, 等. 谷胱甘肽硫转移酶 M5 通过 p38MAPK/NF-κB 途径下调肿瘤坏死因子 α 介导的人支气管上皮细胞炎症水平. 中国呼吸与危护杂志, 2016, 15(2): 171-175.0.
9. Wu X. Transcription start regions in the human genome are favored targets for MLV integration. Science, 2003, 300(5626): 1749-51.
10. Pandya SE, Klimatcheva V. Lentivirus and foamy virus vectors: novel gene therapy tools. Expert Opin Biol Ther, 2001, 1(1): 17-40.
11. Kawakatsu M. Nuclear translocation of glutathione S-transferase p is mediatedby a non-classical localization signal. Biochemical and Biophysical Research Communications: p. 745-750.

Chinese Journal of Respiratory and Critical Care Medicine

GSTM5 nuclear translocation induced by tumor necrosis factor-α

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content