ERK and JNK mediate TNFα-induced p53 activation in apoptotic and autophagic L929 cell death

doi:10.1016/j.bbrc.2008.09.018

Biochemical and Biophysical Research Communications

Volume 376, Issue 3, 21 November 2008, Pages 483-488

https://doi.org/10.1016/j.bbrc.2008.09.018 Get rights and content

Abstract

The object of this study was to investigate the molecular mechanisms mediating TNFα-induced apoptosis and autophagy in L929 cells. Herein, we found that the treatment of L929 cells with TNFα caused a time-dependent increase in p53 activity. The inhibition of p53 activation reduced TNFα-induced apoptosis and autophagy that were accompanied by the decrease in the levels of AIF, Beclin1 and LC3. Subsequently, TNFα activated ERK, JNK and p38 in apoptosis and autopahgy, in which ERK/JNK played a promoting role whereas p38 played an inhibiting one. In addition, TNFα-induced p53 activation was reduced by ERK or JNK inhibition, but it was not affected by p38 inhibition. Further data showed that the inhibition of autophagy reduced TNFα-induced apoptosis in L929 cells. In conclusion, these results demonstrate that TNFα-induced MAPKs mediate p53 activation in apoptotic and autophagic cell death, as well as autophagy may amplify apoptosis when associated with a death signaling pathway.

Section snippets

Materials and methods

Reagents. TNFα was prepared from PMAL-C₂-TNF/JM109 (Escherichia coli) in our laboratory. 3-(4,5-dimetrylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), monodansylcadaverine (MDC), acridine orange (AO), 3-methyladenine (3-MA), p38 inhibitor SB 203580, ERK inhibitor PD 98059, JNK inhibitor SP 600125, and p53 inhibitor pifithrin-α were purchased from Sigma Chemical (St. Louis, MO, USA). Polyclonal antibodies to Beclin 1, LC3, AIF, p38, phospho-p38, ERK, phosphor-ERK,

TNFα-induced apoptotis and autophagy in L929 cells

We examined the effect of TNFα on the growth of L929 cells by the MTT assay. As shown in Fig. 1A, TNFα inhibited L929 cell growth in a time- and dose-dependent manner. To characterize the TNFα-induced L929 cell growth inhibition, we observed the morphologic changes in the cells. When the cells were cultured with 20 ng/ml TNFα for 12 h, the marked apoptotic morphologic alterations, including cell shrinkage, membrane blebbing and nuclear fragmentation were observed (Fig. 1B). For quantitative

Discussion

Of note, the tumor suppressor p53 plays a pivotal role in safeguarding the integrity of the genome and it is also a critical mediator of cell death [13]. TNFα has been reported to induce activation of p53 in various types of cells, suggesting the possibility of involvement of p53 in TNFα-induced cell apoptosis [14], [15]. Consistent with these observations, our results demonstrate that disruption of p53 activation by p53 inhibitor pifithrin-α leads to the decrease in the TNFα-induced cytotoxic

Acknowledgment

We thank Dr. Bo Liu for his critical review on this manuscript.

References (24)

T. Yoshimori
Autophagy: paying charon’s toll
Cell
(2007)
N.A. Maianski et al.
Tumor necrosis factor alpha induces a caspase-independent death pathway in human neutrophils
Blood
(2003)
C. Yu et al.
TMEM74, a lysosome and autophagosome protein, regulates autophagy
Biochem. Biophys. Res. Commun.
(2008)
Y. Cheng et al.
Apoptosis-suppressing and autophagy-promoting effects of calpain on oridonin-induced L929 cell death
Arch. Biochem. Biophys.
(2008)
D. Crighton et al.
DRAM, a p53-induced modulator of autophagy, is critical for apoptosis
Cell
(2006)
S. Lüschen et al.
Inhibition of p38 mitogen-activated protein kinase reduces TNF-induced activation of NF-kappaB, elicits caspase activity, and enhances cytotoxicity
Exp. Cell. Res.
(2004)
K. Matsumaru et al.
Mechanisms for sensitization to TNF-induced apoptosis by acute glutathione depletion in murine hepatocytes
Hepatology
(2003)
L. Harhaji et al.
Aloe emodin inhibits the cytotoxic action of tumor necrosis factor
Eur. J. Pharmacol.
(2007)
L. Moretti et al.
Switch between apoptosis and autophagy: radiation-induced endoplasmic reticulum stress?
Cell Cycle
(2007)
R. Gerl et al.
Apoptosis in the development and treatment of cancer
Carcinogenesis
(2005)

D.J. Klionsky et al.

Autophagy as a regulated pathway of cellular degradation

Science

(2000)

Z.G. Liu et al.

Cellular responses to tumor necrosis factor

Curr. Issues Mol. Biol.

(2001)

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ERK and JNK mediate TNFα-induced p53 activation in apoptotic and autophagic L929 cell death

Abstract

Section snippets

Materials and methods

TNFα-induced apoptotis and autophagy in L929 cells

Discussion

Acknowledgment

Cell

Blood

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

Cell

Exp. Cell. Res.

Hepatology

Eur. J. Pharmacol.

Switch between apoptosis and autophagy: radiation-induced endoplasmic reticulum stress?

Cell Cycle

Apoptosis in the development and treatment of cancer

Carcinogenesis

Autophagy as a regulated pathway of cellular degradation

Science

Cellular responses to tumor necrosis factor

Curr. Issues Mol. Biol.