5-Bromo-2-deoxyuridine activates DNA damage signalling responses and induces a senescence-like phenotype in p16-null lung cancer cells

Masterson, Joanne C.; O’Dea, Shirley

doi:10.1097/cad.0b013e32825209f6

Cited by 36 publications

(33 citation statements)

References 52 publications

(57 reference statements)

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“…We found that cells exposed to the direct DNA-damageinducing agent BrdU induced standard senescence markers, such as p21, fibronectin, IL8 and others, as reported previously (Masterson and O'Dea, 2007;Eriko et al, 1999;Suzuki et al, 2001), and that they are positive for SA-b-gal and show enhanced secretion of IL8 (Cho et al, 2011), which is similar to what is observed in senescent cells generated by other approaches (Coppé et al, 2008). We utilized this unique direct DNA damage approach to activate senescence in cancer cells to determine whether ATM kinase and ROS play distinct roles in the initiation and maintenance of senescence.…”

Section: Discussionsupporting

confidence: 89%

Temporally distinct roles of ATM and ROS in genotoxic-stress-dependent induction and maintenance of cellular senescence

Nair

Bagheri

Saini

2015

Journal of Cell Science

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Cells exposed to genotoxic stress induce cellular senescence through a DNA damage response (DDR) pathway regulated by ATM kinase and reactive oxygen species (ROS). Here, we show that the regulatory roles for ATM kinase and ROS differ during induction and maintenance of cellular senescence. Cells treated with different genotoxic agents were analyzed using specific pathway markers and inhibitors to determine that ATM kinase activation is directly proportional to the dose of the genotoxic stress and that senescence initiation is not dependent on ROS or the p53 status of cells. Cells in which ROS was quenched still activated ATM and initiated the DDR when insulted, and progressed normally to senescence. By contrast, maintenance of a viable senescent state required the presence of ROS as well as activated ATM. Inhibition or removal of either of the components caused cell death in senescent cells, through a deregulated ATM-ROS axis. Overall, our work demonstrates existence of an intricate temporal hierarchy between genotoxic stress, DDR and ROS in cellular senescence. Our model reports the existence of different stages of cellular senescence with distinct regulatory networks.

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Section: Discussionsupporting

confidence: 89%

Temporally distinct roles of ATM and ROS in genotoxic-stress-dependent induction and maintenance of cellular senescence

Nair

Bagheri

Saini

2015

Journal of Cell Science

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“…Pulse-chase analysis demonstrated that CD49f bright /Sca1 1 /ALDH 1 -derived rimmed clones contained rare LRCs and abundant label-diluting cells (LDCs) that were localized to the clone rim ( Figures 3I and 3J). A similar distribution of LRCs and LDCs in the K5/TRE-histone 2B model ( Figure E4) indicated that BrdU toxicity did not affect proliferation of central region cells (32).…”

Section: Basal Cellsmentioning

confidence: 64%

A Single Cell Functions as a Tissue-Specific Stem Cell and theIn VitroNiche-Forming Cell

Ghosh

Helm²,

Smith³

et al. 2011

Am J Respir Cell Mol Biol

102

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Tissue-specific stem cell (TSC) behavior is determined by the stem cell niche. However, delineation of the TSC-niche interaction requires purification of both entities. We reasoned that the niche could be defined by the location of the TSC. We demonstrate that a single CD49f bright /Sca1 1 /ALDH 1 basal cell generates rare label-retaining cells and abundant label-diluting cells. Label-retaining and labeldiluting cells were located in the rimmed domain of a unique clone type, the rimmed clone. The TSC property of self-renewal was tested by serial passage at clonal density and analysis of clone-forming cell frequency. A single clone could be passaged up to five times and formed only rimmed clones. Thus, rimmed clone formation was a cell-intrinsic property. Differentiation potential was evaluated in air-liquid interface cultures. Homogenous cultures of rimmed clones were highly mitotic but were refractory to standard differentiation signals. However, rimmed clones that were cocultured with unfractionated tracheal cells generated each of the cell types found in the tracheal epithelium. Thus, the default niche is promitotic: Multipotential differentiation requires adaptation of the niche. Because lung TSCs are typically evaluated after injury, the behavior of CD49f bright / Sca1 1 /ALDH 1 cells was tested in normal and naphthalene-treated mice. These cells were mitotically active in the normal and repaired epithelium, their proliferation rate increased in response to injury, and they retained label for 34 days. We conclude that the CD49f bright /Sca1 1 /ALDH 1 tracheal basal cell is a TSC, that it generates its own niche in vitro, and that it participates in tracheal epithelial homeostasis and repair.

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“…Since BrdU incorporation during active DNA synthesis occurs during DNA replication and DNA repair [32], and BrdU incorporation can activate DNA damage responses [33], counting BrdU-labeled cells may overestimate the number of mitotic cells, especially when cells are undergoing apoptosis. Here, we detected elevated expression of Ki-67, a marker for cell proliferation, in the DKO but not TKO ileum and colon.…”

Section: Discussionmentioning

confidence: 99%

Nox1 causes ileocolitis in mice deficient in glutathione peroxidase-1 and -2

Esworthy

Kim

Chow

et al. 2014

Free Radical Biology and Medicine

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We previously reported that mice deficient in two Se-dependent glutathione peroxidases, GPx1 and GPx2, have spontaneous ileocolitis. Disease severity depends on mouse genetic background. While C57BL/6J (B6) GPx1/2-DKO mice have moderate ileitis and mild colitis, 129S1Svlm/J (129) DKO mice have severe ileocolitis. Since GPxs are antioxidant enzymes, we hypothesized that elevated reactive oxygen species (ROS) trigger inflammation in these DKO mice. To test whether NADPH oxidase 1 (Nox1) contributes to colitis, we generated B6 triple-KO (TKO) mice to study their phenotype. Since the Nox1 gene is X-linked, we analyzed the effect of NOX1 on male B6 TKO and female B6 DKO with the Nox1+/− (het-TKO) genotype. We found that the male TKO and female het-TKO mice are virtually disease-free when monitored from 8 through 50 days of age. Male TKO and female het-TKO mice have nearly no signs of disease (e.g. lethargy and perianal alopecia) that is often exhibited in the DKO mice; further, the slower growth rate of DKO mice is almost completely eliminated in maleTKO and female het-TKO mice. Male TKO and female het-TKO mice no longer have the shortened small intestine present in the DKO mice. Finally, the pathological characteristics of the DKO ileum, including the high number of crypt apoptosis (analyzed by apoptotic figures, TUNEL and cleaved caspase-3 immunohistochemical staining), and high numbers of Ki-67-positive crypt epithelium cells and elevated levels of monocytes expressing myeloperoxidase are all significantly decreased in male TKO mice. The attenuated ileal and colonic pathology is also evident in female het-DKO mice. Furthermore, the male DKO ileum has 8-fold higher TNF cytokine levels than TKO ileum. Nox1 mRNA is highly elevated in both B6 and 129 DKO ileum compared to that in WT mouse ileum. Taken together, we propose that ileocolitis in the DKO mice is caused by NOX1, which is induced by TNF. The milder disease in female het-TKO intestine is likely due to random or imprinted X-chromosome inactivation, which produces mosaic Nox1 expression.

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5-Bromo-2-deoxyuridine activates DNA damage signalling responses and induces a senescence-like phenotype in p16-null lung cancer cells

Cited by 36 publications

References 52 publications

Temporally distinct roles of ATM and ROS in genotoxic-stress-dependent induction and maintenance of cellular senescence

Temporally distinct roles of ATM and ROS in genotoxic-stress-dependent induction and maintenance of cellular senescence

A Single Cell Functions as a Tissue-Specific Stem Cell and theIn VitroNiche-Forming Cell

Nox1 causes ileocolitis in mice deficient in glutathione peroxidase-1 and -2

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