Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Molecular Cancer
Published in: Molecular Cancer 1/2010

Open Access 01-12-2010 | Research

Selective unresponsiveness to the inhibition of p38 MAPK activation by cAMP helps L929 fibroblastoma cells escape TNF-α-induced cell death

Authors: Jing Wang, Ruihong Tang, Ming Lv, Jiyan Zhang, Beifen Shen

Published in: Molecular Cancer | Issue 1/2010

Login to get access

Abstract

Background

The cyclic AMP (cAMP) signaling pathway has been reported to either promote or suppress cell death, in a cell context-dependent manner. Our previous study has shown that the induction of dynein light chain (DLC) by cAMP response element-binding protein (CREB) is required for cAMP-mediated inhibition of mitogen-activated protein kinase (MAPK) p38 activation in fibroblasts, which leads to suppression of NF-κB activity and promotion of tumor necrosis factor-α (TNF-α)-induced cell death. However, it remains unknown whether this regulation is also applicable to fibroblastoma cells.

Methods

Intracellular cAMP was determined in L929 fibroblastoma cells after treatment of the cells with various cAMP elevation agents. Effects of cAMP in the presence or absence of the RNA synthesis inhibitor actinomycin D or small interfering RNAs (siRNAs) against CREB on TNF-α-induced cell death in L929 cells were measured by propidium iodide (PI) staining and subsequent flow cytomety. The activation of p38 and c-Jun N-terminal protein kinase (JNK), another member of MAPK superfamily, was analyzed by immunoblotting. JNK selective inhibitor D-JNKi1 and p38 selective inhibitor SB203580 were included to examine the roles of JNK and p38 in this process. The expression of DLC or other mediators of cAMP was analyzed by immunoblotting. After ectopic expression of DLC with a transfection marker GFP, effects of cAMP on TNF-α-induced cell death in GFP+ cells were measured by PI staining and subsequent flow cytomety.

Results

Elevation of cAMP suppressed TNF-α-induced necrotic cell death in L929 fibroblastoma cells via CREB-mediated transcription. The pro-survival role of cAMP was associated with selective unresponsiveness of L929 cells to the inhibition of p38 activation by cAMP, even though cAMP significantly inhibited the activation of JNK under the same conditions. Further exploration revealed that the induction of DLC, the major mediator of p38 inhibition by cAMP, was impaired in L929 cells. Enforced inhibition of p38 activation by using p38 specific inhibitor or ectopic expression of DLC reversed the protection of L929 cells by cAMP from TNF-α-induced cell death.

Conclusion

These data suggest that the lack of a pro-apoptotic pathway in tumor cells leads to a net survival effect of cAMP.
Appendix
Available only for authorised users
Literature
1.
Rosenberg D, Groussin L, Jullian E, Perlemoine K, Bertagna X, Bertherat J: Role of the PKA-regulated transcription factor CREB in development and tumorigenesis of endocrine tissues. Ann N Y Acad Sci. 2002, 968: 65-74. 10.1111/j.1749-6632.2002.tb04327.xCrossRefPubMed
2.
James MA, Lu Y, Liu Y, Vikis HG, You M: RGS17, an overexpressed gene in human lung and prostate cancer, induces tumor cell proliferation through the cyclic AMP-PKA-CREB pathway. Cancer Res. 2009, 69: 2108-2116. 10.1158/0008-5472.CAN-08-3495PubMedCentralCrossRefPubMed
3.
Sastry KS, Karpova Y, Prokopovich S, Smith AJ, Essau B, Gersappe A, Carson JP, Weber MJ, Register TC, Chen YQ: Epinephrine protects cancer cells from apoptosis via activation of cAMP-dependent protein kinase and BAD phosphorylation. J Biol Chem. 2007, 282: 14094-14100. 10.1074/jbc.M611370200CrossRefPubMed
4.
Garcia-Bermejo L, Perez C, Vilaboa NE, de Blas E, Aller P: cAMP increasing agents attenuate the generation of apoptosis by etoposide in promonocytic leukemia cells. J Cell Sci. 1998, 111 (Pt 5): 637-644.PubMed
5.
von Knethen A, Brune B: Attenuation of macrophage apoptosis by the cAMP-signaling system. Mol Cell Biochem. 2000, 212: 35-43. 10.1023/A:1007124203607CrossRefPubMed
6.
Boucher MJ, Duchesne C, Laine J, Morisset J, Rivard N: cAMP protection of pancreatic cancer cells against apoptosis induced by ERK inhibition. Biochem Biophys Res Commun. 2001, 285: 207-216. 10.1006/bbrc.2001.5147CrossRefPubMed
7.
Chiu HF, Chih TT, Hsian YM, Tseng CH, Wu MJ, Wu YC: Bullatacin, a potent antitumor Annonaceous acetogenin, induces apoptosis through a reduction of intracellular cAMP and cGMP levels in human hepatoma 2.2.15 cells. Biochem Pharmacol. 2003, 65: 319-327. 10.1016/S0006-2952(02)01554-XCrossRefPubMed
8.
Nishihara H, Hwang M, Kizaka-Kondoh S, Eckmann L, Insel PA: Cyclic AMP promotes cAMP-responsive element-binding protein-dependent induction of cellular inhibitor of apoptosis protein-2 and suppresses apoptosis of colon cancer cells through ERK1/2 and p38 MAPK. J Biol Chem. 2004, 279: 26176-26183. 10.1074/jbc.M313346200CrossRefPubMed
9.
Sakamoto KM, Frank DA: CREB in the pathophysiology of cancer: implications for targeting transcription factors for cancer therapy. Clin Cancer Res. 2009, 15: 2583-2587. 10.1158/1078-0432.CCR-08-1137PubMedCentralCrossRefPubMed
10.
Naderi EH, Findley HW, Ruud E, Blomhoff HK, Naderi S: Activation of cAMP signaling inhibits DNA damage-induced apoptosis in BCP-ALL cells through abrogation of p53 accumulation. Blood. 2009, 114: 608-618. 10.1182/blood-2009-02-204883CrossRefPubMed
11.
Guevara Patino JA, Ivanov VN, Lacy E, Elkon KB, Marino MW, Nikolic-Zugic J: TNF-alpha is the critical mediator of the cyclic AMP-induced apoptosis of CD8+4+ double-positive thymocytes. J Immunol. 2000, 164: 1689-1694.CrossRefPubMed
12.
Kostylina G, Simon D, Fey MF, Yousefi S, Simon HU: Neutrophil apoptosis mediated by nicotinic acid receptors (GPR109A). Cell Death Differ. 2008, 15: 134-142. 10.1038/sj.cdd.4402238CrossRefPubMed
13.
Negrotto S, Pacienza N, D'Atri LP, Pozner RG, Malaver E, Torres O, Lazzari MA, Gomez RM, Schattner M: Activation of cyclic AMP pathway prevents CD34(+) cell apoptosis. Exp Hematol. 2006, 34: 1420-1428. 10.1016/j.exphem.2006.05.017CrossRefPubMed
14.
Zhang J, Bui TN, Xiang J, Lin A: Cyclic AMP inhibits p38 activation via CREB-induced dynein light chain. Mol Cell Biol. 2006, 26: 1223-1234. 10.1128/MCB.26.4.1223-1234.2006PubMedCentralCrossRefPubMed
15.
Zhang J, Wang Q, Zhu N, Yu M, Shen B, Xiang J, Lin A: Cyclic AMP inhibits JNK activation by CREB-mediated induction of c-FLIP(L) and MKP-1, thereby antagonizing UV-induced apoptosis. Cell Death Differ. 2008, 15: 1654-1662. 10.1038/cdd.2008.87CrossRefPubMed
16.
Hibi M, Lin A, Smeal T, Minden A, Karin M: Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev. 1993, 7: 2135-2148. 10.1101/gad.7.11.2135CrossRefPubMed
17.
Han J, Lee JD, Bibbs L, Ulevitch RJ: A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. Science. 1994, 265: 808-811. 10.1126/science.7914033CrossRefPubMed
18.
Rouse J, Cohen P, Trigon S, Morange M, Alonso-Llamazares A, Zamanillo D, Hunt T, Nebreda AR: A novel kinase cascade triggered by stress and heat shock that stimulates MAPKAP kinase-2 and phosphorylation of the small heat shock proteins. Cell. 1994, 78: 1027-1037. 10.1016/0092-8674(94)90277-1CrossRefPubMed
19.
Chang L, Karin M: Mammalian MAP kinase signalling cascades. Nature. 2001, 410: 37-40. 10.1038/35065000CrossRefPubMed
20.
Lin A: Activation of the JNK signaling pathway: breaking the brake on apoptosis. Bioessays. 2003, 25: 17-24. 10.1002/bies.10204CrossRefPubMed
21.
Zarubin T, Han J: Activation and signaling of the p38 MAP kinase pathway. Cell Res. 2005, 15: 11-18. 10.1038/sj.cr.7290257CrossRefPubMed
22.
Zhang J, Shen B, Lin A: Novel strategies for inhibition of the p38 MAPK pathway. Trends Pharmacol Sci. 2007, 28: 286-295. 10.1016/j.tips.2007.04.008CrossRefPubMed
23.
Chang L, Kamata H, Solinas G, Luo JL, Maeda S, Venuprasad K, Liu YC, Karin M: The E3 ubiquitin ligase itch couples JNK activation to TNFalpha-induced cell death by inducing c-FLIP(L) turnover. Cell. 2006, 124: 601-613. 10.1016/j.cell.2006.01.021CrossRefPubMed
24.
Guo YL, Baysal K, Kang B, Yang LJ, Williamson JR: Correlation between sustained c-Jun N-terminal protein kinase activation and apoptosis induced by tumor necrosis factor-alpha in rat mesangial cells. J Biol Chem. 1998, 273: 4027-4034. 10.1074/jbc.273.7.4027CrossRefPubMed
25.
Ventura JJ, Hubner A, Zhang C, Flavell RA, Shokat KM, Davis RJ: Chemical genetic analysis of the time course of signal transduction by JNK. Mol Cell. 2006, 21: 701-710. 10.1016/j.molcel.2006.01.018CrossRefPubMed
26.
Zhang J, Liu J, Yu C, Lin A: BAD Ser128 is not phosphorylated by c-Jun NH2-terminal kinase for promoting apoptosis. Cancer Res. 2005, 65: 8372-8378. 10.1158/0008-5472.CAN-05-0576CrossRefPubMed
27.
Seamon KB, Padgett W, Daly JW: Forskolin: unique diterpene activator of adenylate cyclase in membranes and in intact cells. Proc Natl Acad Sci USA. 1981, 78: 3363-3367. 10.1073/pnas.78.6.3363PubMedCentralCrossRefPubMed
28.
Fiers W, Beyaert R, Declercq W, Vandenabeele P: More than one way to die: apoptosis, necrosis and reactive oxygen damage. Oncogene. 1999, 18: 7719-7730. 10.1038/sj.onc.1203249CrossRefPubMed
29.
Xie C, Zhang N, Zhou H, Li J, Li Q, Zarubin T, Lin SC, Han J: Distinct roles of basal steady-state and induced H-ferritin in tumor necrosis factor-induced death in L929 cells. Mol Cell Biol. 2005, 25: 6673-6681. 10.1128/MCB.25.15.6673-6681.2005PubMedCentralCrossRefPubMed
30.
Jackisch R, Bruckner-Schmidt R, Hertting G: Endogenously formed PGE2 mediates the increase in cAMP accumulation in rabbit splenic fibroblasts following alpha-receptor stimulation. Eur J Pharmacol. 1981, 71: 151-155. 10.1016/0014-2999(81)90400-3CrossRefPubMed
31.
Mayr B, Montminy M: Transcriptional regulation by the phosphorylation-dependent factor CREB. Nat Rev Mol Cell Biol. 2001, 2: 599-609. 10.1038/35085068CrossRefPubMed
32.
Impey S, McCorkle SR, Cha-Molstad H, Dwyer JM, Yochum GS, Boss JM, McWeeney S, Dunn JJ, Mandel G, Goodman RH: Defining the CREB regulon: a genome-wide analysis of transcription factor regulatory regions. Cell. 2004, 119: 1041-1054.PubMed
33.
Zhang X, Odom DT, Koo SH, Conkright MD, Canettieri G, Best J, Chen H, Jenner R, Herbolsheimer E, Jacobsen E: Genome-wide analysis of cAMP-response element binding protein occupancy, phosphorylation, and target gene activation in human tissues. Proc Natl Acad Sci USA. 2005, 102: 4459-4464. 10.1073/pnas.0501076102PubMedCentralCrossRefPubMed
34.
Binder C, Binder L, Kroemker M, Schulz M, Hiddemann W: Influence of cycloheximide-mediated downregulation of glucose transport on TNF alpha-induced apoptosis. Exp Cell Res. 1997, 236: 223-230. 10.1006/excr.1997.3718CrossRefPubMed
35.
Wang L, Du F, Wang X: TNF-alpha induces two distinct caspase-8 activation pathways. Cell. 2008, 133: 693-703. 10.1016/j.cell.2008.03.036CrossRefPubMed
36.
Ahn S, Olive M, Aggarwal S, Krylov D, Ginty DD, Vinson C: A dominant-negative inhibitor of CREB reveals that it is a general mediator of stimulus-dependent transcription of c-fos. Mol Cell Biol. 1998, 18: 967-977.PubMedCentralCrossRefPubMed
37.
Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C: A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nat Med. 2003, 9: 1180-1186. 10.1038/nm911CrossRefPubMed
38.
Cuenda A, Rouse J, Doza YN, Meier R, Cohen P, Gallagher TF, Young PR, Lee JC: SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett. 1995, 364: 229-233. 10.1016/0014-5793(95)00357-FCrossRefPubMed
39.
Whitmarsh AJ, Yang SH, Su MS, Sharrocks AD, Davis RJ: Role of p38 and JNK mitogen-activated protein kinases in the activation of ternary complex factors. Mol Cell Biol. 1997, 17: 2360-2371.PubMedCentralCrossRefPubMed
40.
Kunisch E, Jansen A, Kojima F, Loffler I, Kapoor M, Kawai S, Rubio I, Crofford LJ, Kinne RW: Prostaglandin E2 differentially modulates proinflammatory/prodestructive effects of TNF-alpha on synovial fibroblasts via specific E prostanoid receptors/cAMP. J Immunol. 2009, 183: 1328-1336. 10.4049/jimmunol.0900801CrossRefPubMed
41.
Irmler M, Thome M, Hahne M, Schneider P, Hofmann K, Steiner V, Bodmer JL, Schroter M, Burns K, Mattmann C: Inhibition of death receptor signals by cellular FLIP. Nature. 1997, 388: 190-195. 10.1038/40657CrossRefPubMed
42.
Kim YH, Kim SS: Increase of MnSOD expression and decrease of JNK activity determine the TNF sensitivity in bcl2-transfected L929 cells. Cytokine. 1999, 11: 274-281. 10.1006/cyto.1998.0414CrossRefPubMed
43.
Luschen S, Scherer G, Ussat S, Ungefroren H, Adam-Klages S: Inhibition of p38 mitogen-activated protein kinase reduces TNF-induced activation of NF-kappaB, elicits caspase activity, and enhances cytotoxicity. Exp Cell Res. 2004, 293: 196-206. 10.1016/j.yexcr.2003.10.009CrossRefPubMed
Metadata
Title
Selective unresponsiveness to the inhibition of p38 MAPK activation by cAMP helps L929 fibroblastoma cells escape TNF-α-induced cell death
Authors
Jing Wang
Ruihong Tang
Ming Lv
Jiyan Zhang
Beifen Shen
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2010
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-9-6

Other articles of this Issue 1/2010

Molecular Cancer 1/2010 Go to the issue

Research

The two most common histological subtypes of malignant germ cell tumour are distinguished by global microRNA profiles, associated with differential transcription factor expression

Research

Sulforaphane induces cell cycle arrest by protecting RB-E2F-1 complex in epithelial ovarian cancer cells

Research

c-Jun NH2-terminal kinase-dependent upregulation of DR5 mediates cooperative induction of apoptosis by perifosine and TRAIL

Research

Reptin is required for the transcription of telomerase reverse transcriptase and over-expressed in gastric cancer

Research

Prognostic significance and therapeutic implications of centromere protein F expression in human nasopharyngeal carcinoma

Review

Survival signalling and apoptosis resistance in glioblastomas: opportunities for targeted therapeutics
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits