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Tumor necrosis factor-alpha induces MMP-9 expression via p42/p44 MAPK, JNK, and nuclear factor-kappaB in A549 cells.
Toxicol Appl Pharmacol. 2008 Jun 15; 229(3):386-98.TA

Abstract

Matrix metalloproteinases (MMPs), in particular MMP-9, have been shown to be induced by cytokines including tumor necrosis factor-alpha (TNF-alpha) and contributes to airway inflammation. However, the mechanisms underlying MMP-9 expression induced by TNF-alpha in human A549 cells remain unclear. Here, we showed that TNF-alpha induced production of MMP-9 protein and mRNA is determined by zymographic, Western blotting, RT-PCR and ELISA assay, which were attenuated by inhibitors of MEK1/2 (U0126), JNK (SP600125), and NF-kappaB (helenalin), and transfection with dominant negative mutants of ERK2 (DeltaERK) and JNK (DeltaJNK), and siRNAs for MEK1, p42 and JNK2. TNF-alpha-stimulated phosphorylation of p42/p44 MAPK and JNK were attenuated by pretreatment with the inhibitors U0126 and SP600125 or transfection with dominant negative mutants of DeltaERK and DeltaJNK. Furthermore, the involvement of NF-kappaB in TNF-alpha-induced MMP-9 production was consistent with that TNF-alpha-stimulated degradation of IkappaB-alpha and translocation of NF-kappaB into the nucleus which were blocked by helenalin, but not by U0126 and SP600125, revealed by immunofluorescence staining. The regulation of MMP-9 gene transcription by MAPKs and NF-kappaB was further confirmed by gene luciferase activity assay. MMP-9 promoter activity was enhanced by TNF-alpha in A549 cells transfected with wild-type MMP-9-Luc, which was inhibited by helenalin, U0126, or SP600125. In contrast, TNF-alpha-stimulated MMP-9 luciferase activity was totally lost in cells transfected with mutant-NF-kappaB MMP-9-luc. Moreover, pretreatment with actinomycin D and cycloheximide attenuated TNF-alpha-induced MMP-9 expression. These results suggest that in A549 cells, phosphorylation of p42/p44 MAPK, JNK, and transactivation of NF-kappaB are essential for TNF-alpha-induced MMP-9 gene expression.

Authors+Show Affiliations

Department of Anesthetics, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18336852

Citation

Lin, Chih-Chung, et al. "Tumor Necrosis Factor-alpha Induces MMP-9 Expression Via P42/p44 MAPK, JNK, and Nuclear factor-kappaB in A549 Cells." Toxicology and Applied Pharmacology, vol. 229, no. 3, 2008, pp. 386-98.
Lin CC, Tseng HW, Hsieh HL, et al. Tumor necrosis factor-alpha induces MMP-9 expression via p42/p44 MAPK, JNK, and nuclear factor-kappaB in A549 cells. Toxicol Appl Pharmacol. 2008;229(3):386-98.
Lin, C. C., Tseng, H. W., Hsieh, H. L., Lee, C. W., Wu, C. Y., Cheng, C. Y., & Yang, C. M. (2008). Tumor necrosis factor-alpha induces MMP-9 expression via p42/p44 MAPK, JNK, and nuclear factor-kappaB in A549 cells. Toxicology and Applied Pharmacology, 229(3), 386-98. https://doi.org/10.1016/j.taap.2008.01.032
Lin CC, et al. Tumor Necrosis Factor-alpha Induces MMP-9 Expression Via P42/p44 MAPK, JNK, and Nuclear factor-kappaB in A549 Cells. Toxicol Appl Pharmacol. 2008 Jun 15;229(3):386-98. PubMed PMID: 18336852.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Tumor necrosis factor-alpha induces MMP-9 expression via p42/p44 MAPK, JNK, and nuclear factor-kappaB in A549 cells. AU - Lin,Chih-Chung, AU - Tseng,Hsiao-Wei, AU - Hsieh,Hsi-Lung, AU - Lee,Chiang-Wen, AU - Wu,Cheng-Ying, AU - Cheng,Ching-Yi, AU - Yang,Chuen-Mao, Y1 - 2008/02/09/ PY - 2007/11/01/received PY - 2008/01/15/revised PY - 2008/01/26/accepted PY - 2008/3/14/pubmed PY - 2008/7/22/medline PY - 2008/3/14/entrez SP - 386 EP - 98 JF - Toxicology and applied pharmacology JO - Toxicol. Appl. Pharmacol. VL - 229 IS - 3 N2 - Matrix metalloproteinases (MMPs), in particular MMP-9, have been shown to be induced by cytokines including tumor necrosis factor-alpha (TNF-alpha) and contributes to airway inflammation. However, the mechanisms underlying MMP-9 expression induced by TNF-alpha in human A549 cells remain unclear. Here, we showed that TNF-alpha induced production of MMP-9 protein and mRNA is determined by zymographic, Western blotting, RT-PCR and ELISA assay, which were attenuated by inhibitors of MEK1/2 (U0126), JNK (SP600125), and NF-kappaB (helenalin), and transfection with dominant negative mutants of ERK2 (DeltaERK) and JNK (DeltaJNK), and siRNAs for MEK1, p42 and JNK2. TNF-alpha-stimulated phosphorylation of p42/p44 MAPK and JNK were attenuated by pretreatment with the inhibitors U0126 and SP600125 or transfection with dominant negative mutants of DeltaERK and DeltaJNK. Furthermore, the involvement of NF-kappaB in TNF-alpha-induced MMP-9 production was consistent with that TNF-alpha-stimulated degradation of IkappaB-alpha and translocation of NF-kappaB into the nucleus which were blocked by helenalin, but not by U0126 and SP600125, revealed by immunofluorescence staining. The regulation of MMP-9 gene transcription by MAPKs and NF-kappaB was further confirmed by gene luciferase activity assay. MMP-9 promoter activity was enhanced by TNF-alpha in A549 cells transfected with wild-type MMP-9-Luc, which was inhibited by helenalin, U0126, or SP600125. In contrast, TNF-alpha-stimulated MMP-9 luciferase activity was totally lost in cells transfected with mutant-NF-kappaB MMP-9-luc. Moreover, pretreatment with actinomycin D and cycloheximide attenuated TNF-alpha-induced MMP-9 expression. These results suggest that in A549 cells, phosphorylation of p42/p44 MAPK, JNK, and transactivation of NF-kappaB are essential for TNF-alpha-induced MMP-9 gene expression. SN - 0041-008X UR - https://www.unboundmedicine.com/medline/citation/18336852/Tumor_necrosis_factor_alpha_induces_MMP_9_expression_via_p42/p44_MAPK_JNK_and_nuclear_factor_kappaB_in_A549_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0041-008X(08)00062-8 DB - PRIME DP - Unbound Medicine ER -