Checkpoint kinase (Chk1) and reactive oxygen species induced oncotic cell death during the cell cycle in LLC-PK1 cells

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Research Paper 01/06/2012
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Checkpoint kinase (Chk1) and reactive oxygen species induced oncotic cell death during the cell cycle in LLC-PK1 cells

Hashimul Ehsan
Int. J. Biosci.2( 6), 24-31, June 2012.
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2,3,5–tris-(Glutathion-S-yl)hydroquinone (TGHQ) is a reactive oxygen species (ROS) generating metabolite of the nephotoxicant, hydroquinone. In renal proximal tubule epithelial cells (LLC-PK1), TGHQ catalyzes the formation of single strand breaks in DNA, followed by growth arrest and oncotic/necrotic death during the cell cycle. Treatment of synchronized LLC-PK1 cells with TGHQ causes an initial decrease in the expression of the DNA damage inducible protein kinase chk1. The degradation of chk1 in highly synchronized LLC-PK1 cells might induce mitotic catastrophe, a potential mechanism of cell death in this model.


Abraham RT. 2001. Cell cycle checkpoint signaling through the ATM and ATR kinases, Genes Dev. 15, 2177–2196.

Boddy MN, B. Furnari O, Mondesert B, Russell P 1998. Replication checkpoint enforced by kinases Cds1 and Chk1. Science 280, 909-912.

Cortez D, Guntuku S, Qin J, Elledge SJ. 2001. ATR  and  ATRIP:  partners  in  checkpoint  signaling. Science 5547, 1713-6.

Ehsan H, Reichheld JP, Roef L, Witters E, Lardon F, Van Bockstaele D, Van Montagu M, Inze D, Van Onckelen H. 1998. Effect of indomethacin on cell cycle dependent cyclic AMP fluxes in tobacco BY-2 cells. FEBS Letters 422(2), 165-9.

Francesconi SM, Grenon D, Bouvier C, Baldacci G. 1997. p56chk1 protein kinase is required for the DNA replication checkpoint at 37oC in fission yeast. EMBO J. 16, 1332-1341.

Jeong JK, Dybing E, Soderlund E, Brunborg G, Holme JA, Lau SS, Monks TJ. 1997a. DNA damage, gadd153 expression, and cytotoxicity in plateau-phase renal proximal tubular epithelial cells treated with a quinol thioether. Arch Biochem Biophys. 341, 300-308.

Jeong JK, Huang Q, Lau S.S, Monks TJ. 1997b. The response of renal tubular epithelial cells to physiologically and chemically induced growth arrest. J Biol Chem 272, 7511-7518.

Jeong JK, Wogan GN, Lau SS, Monks TJ. 1999. Quinol-glutathione conjugate-induced mutation spectra in the supF gene replicated in human AD293 cells and bacterial MBL50 cells. Cancer Res. 15, 3641-3645.

Lindsay H D, Griffiths DJ, Edwards RJ, Christensen PU, Murray JM, Osman F, Walworth N, Carr AM. 1998. S-phase-specific activation of Cds1 kinase defines a subpathway of the checkpoint response in Schizosaccharomyces pombe. Genes Dev. 12, 382-395.

Lu X, Nannenga B, Donehower LA. 2005. PPM1D dephosphorylates Chk1 and p53 and abrogates cell cycle checkpoints. Genes Dev. 19, 1162–1174.

Mertens JJ, Gibson NW, Lau SS, Monks TJ. 1995.  Reactive oxygen species and DNA damage in 2- bromo-(glutathion-S-yl) hydroquinone-mediated cytotoxicity. Arch Biochem Biophys. 320, 51-58.

Monks TJ, Lau SS. 1998. The pharmacology and toxicology of polyphenolic-glutathione conjugates. Annu Rev Pharmacol Toxicol. 38, 229-255.

Morgan MA, Parsels LA, Parsels JD, Lawerence TS, Maybaum J. 2006. The relationship of premature mitosis to cytotoxicity in response to checkpoint abrogation and antimetabolite treatment. Cell Cycle 5:17, 1-6.

Niida H, Tsuge S, Katsuno Y, Konishi A, Takeda N. Nakanishi M. 2005. Depletion of Chk1 leads to premature activation of Cdc2-cyclin B and mitotic catastrophy. J. Biol. Chem. 280 (47), 39246-52.

Nurse P. 2000. A long twentieth century of the cell cycle and beyond. Cell 100, 71-78.

Ramachandiran S, Huang Q, Dong J, Lau SS, Monks TJ. 2002. Mitogen-activated protein kinases contribute to reactive oxygen species-induced cell death in renal proximal tubule epithelial cells. Chem. Res. Toxicol. 12, 1635-1642.

Tikoo K, Lau SS, Monks TJ. 2001. Histone H3 phosphorylation is coupled to poly-(ADP-ribosylation) during reactive oxygen species-induced cell death in renal proximal tubular epithelial cells. Mol Pharmacol. 60, 394-402.

Walworth N C, Bernards R. 1996. Rad-dependent response of the chk1-encoded protein kinase at the DNA damage checkpoint. Science 271, 353-356.

Yoon HS, Monks TJ, Everitt JI, Walker CL, Lau SS. 2002. Cell proliferation is insufficient, but loss of tuberin is necessary, for chemically induced nephrocarcinogenicity. Am J Physiol Renal Physiol. 283, 262-270.

Zeng YKC, Forbes Z, Wu S, Moreno H, Piwnica-Worms, Enoch T. 1998. Replication checkpoint requires phosphorylation of the phosphatase Cdc25 by Cds1 or Chk1. Nature 395, 507-510.

Zhang YW, Otternes DM, Chiang, GG, Xie W, Liu Y, Mercurio F, Abraham RT. 2005. Genotoxic stress targets human Chk1 for degradation by the ubiquitin-proteasome pathway. Molecular Cell 19 (5), 607-618.

Zhou BB, Elledge SJ. 2000. The DNA damage response: putting checkpoints in perspective. Nature 408, 433-439.

Zou L, Elledge SJ. 2003. Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300, 1542-1548.