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J. Biol. Chem., Vol. 280, Issue 28, 26193-26199, July 15, 2005

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Chloramphenicol-induced Mitochondrial Stress Increases p21 Expression and Prevents Cell Apoptosis through a p21-dependent Pathway^*

Ching-Hao Li, Su-Liang Tzeng, Yu-Wen Cheng, and Jaw-Jou Kang

From the Institute of Toxicology, College of Medicine, National Taiwan University, Taipei 100, Taiwan

Pretreatment of HepG2 and H1299 cells with chloramphenicol rendered the cells resistant to mitomycin-induced apoptosis. Both mitomycin-induced caspase 3 activity and PARP activation were also inhibited. The mitochondrial DNA-encoded Cox I protein, but not nuclear-encoded proteins, was down-regulated in chloramphenicol-treated cells. Cellular levels of the p21^waf1/cip1 protein and p21^waf1/cip1 mRNA were increased through a p53-independent pathway, possibly because of the stabilization of p21^waf1/cip1 mRNA in chloramphenicol-treated cells. The p21^waf1/cip1 was redistributed from the perinuclear region to the cytoplasm and co-localized with mitochondrial marker protein. Several morphological changes and activation of the senescence-associated biomarker, SA -galactosidase, were observed in these cells. Both p21^waf1/cip1 antisense and small interfering RNA could restore apoptotic-associated caspase 3 activity, PARP activation, and sensitivity to mitomycin-induced apoptosis. Similar effects were seen with other antibiotics that inhibit mitochondrial translation, including minocycline, doxycycline, and clindamycin. These findings suggested that mitochondrial stress causes resistance to apoptosis through a p21-dependent pathway.

Received for publication, February 4, 2005 , and in revised form, April 11, 2005.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Institute of Toxicology, College of Medicine, National Taiwan University, No. 1 Jen-Ai Rd., Section 1, Taipei 100, Taiwan. Tel.: 886-2-23123456 (ext. 8603); Fax: 886-2-23410217; E-mail: jjkang{at}ha.mc.ntu.edu.tw.

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