DNA damage can impair normal cellular functions and result in various pathophysiological processes including cardiovascular diseases and cancer. We compared the genotoxic potential of diverse DNA damaging agents, and focused on their effects on the DNA damage response (DDR) and cell fate in human lung cells BEAS-2B. Polycyclic aromatic hydrocarbons [PAHs; benzo[a]pyrene (B[a]P), 1-nitropyrene (1-NP)] induced DNA strand breaks and oxidative damage to DNA; anticancer drugs doxorubicin (DOX) and 5-bromo-2'-deoxyuridine (BrdU) were less effective. DOX triggered the most robust p53 signaling indicating activation of DDR, followed by cell cycle arrest in the G2/M phase, induction of apoptosis and senescence, possibly due to the severe and irreparable DNA lesions. BrdU not only activated p53, but also increased the percentage of G1-phased cells and caused a massive accumulation of senescent cells. In contrast, regardless the activation of p53, both PAHs did not substantially affect the cell cycle distribution or senescence. Finally, a small fraction of cells accumulated only in the G2/M phase and exhibited increased cell death after the prolonged incubation with B[a]P. Overall, we characterized differential responses to diverse DNA damaging agents resulting in specific cell fate and highlighted the key role of DNA lesion type and the p53 signaling persistence.
Cellular senescence guards against cancer and modulates aging; however, the underlying mechanisms remain poorly understood. Here, we show that genotoxic drugs capable of inducing premature senescence in normal and cancer cells, such as 5-bromo-2'-deoxyuridine (BrdU), distamycin A (DMA), aphidicolin and hydroxyurea, persistently activate Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling and expression of interferon-stimulated genes (ISGs), such as MX1, OAS, ISG15, STAT1, PML, IRF1 and IRF7, in several human cancer cell lines. JAK1/STAT-activating ligands, interleukin 10 (IL10), IL20, IL24, interferon gamma (IFNgamma), IFNbeta and IL6, were also expressed by senescent cells, supporting autocrine/paracrine activation of JAK1/STAT. Furthermore, cytokine genes, including proinflammatory IL1, tumor necrosis factor and transforming growth factor families, were highly expressed. The strongest inducer of JAK/STAT signaling, cytokine production and senescence was BrdU combined with DMA. RNA interference-mediated knockdown of JAK1 abolished expression of ISGs, but not DNA damage signaling or senescence. Thus, although DNA damage signaling, p53 and RB activation, and the cytokine/chemokine secretory phenotype are apparently shared by all types of senescence, our data reveal so far unprecedented activation of the IFNbeta-STAT1-ISGs axis, and indicate a less prominent causative role of IL6-JAK/STAT signaling in genotoxic drug-induced senescence compared with reports on oncogene-induced or replicative senescence. These results highlight shared and unique features of drug-induced cellular senescence, and implicate induction of cancer secretory phenotype in chemotherapy.
- MeSH
- bromodeoxyuridin farmakologie MeSH
- cytokiny genetika metabolismus MeSH
- distamyciny farmakologie MeSH
- HeLa buňky MeSH
- interferony genetika metabolismus MeSH
- interleukin-10 genetika metabolismus MeSH
- interleukin-6 genetika metabolismus MeSH
- interleukin-8 genetika metabolismus MeSH
- interleukiny genetika metabolismus MeSH
- Janus kinasa 1 genetika metabolismus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- RNA interference MeSH
- signální transdukce účinky léků MeSH
- stárnutí buněk účinky léků MeSH
- synergismus léků MeSH
- transkripční faktor STAT1 genetika metabolismus MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
The mechanisms of sodium selenite-induced cell death in cervical carcinoma cells were studied during 24 h of exposure in the HeLa Hep-2 cell line. Selenite at the employed concentrations of 5 and 50 micromol/L produced time- and dose-dependent suppression of DNA synthesis and induced DNA damage which resulted in phosphorylation of histone H2A.X. These effects were influenced by pretreatment of cells with the SOD/catalase mimetic MnTMPyP or glutathione-depleting buthionine sulfoximine, suggesting the significant role of selenite-generated oxidative stress. Following the DNA damage, selenite activated p53-dependent pathway as evidenced by the appearance of phosphorylated p53 and accumulation of p21 in the treated cells. Concomitantly, selenite activated p38 pathway but its effect on JNK was very weak. p53- and p38-dependent signaling led to the accumulation of Bax protein, which was preventable by specific inhibitors of p38 (SB 203580) and p53 (Pifithrin-alpha). Mitochondria in selenite-treated cells changed their dynamics (shape and localization) and released AIF and Smac/Diablo, which initiated caspase-independent apoptosis as confirmed by the caspase-3 activity assay and the low effect of caspase inhibitors z-DEVD-fmk and z-VAD-fmk on cell death. We conclude that selenite induces caspase-independent apoptosis in cervical carcinoma cells mostly by oxidative stress-mediated activation of p53 and p38 pathways, but other selenite-mediated effects, in particular mitochondria-specific ones, are also involved.
- MeSH
- aktivace enzymů účinky léků MeSH
- bromodeoxyuridin farmakologie metabolismus MeSH
- buněčná smrt účinky léků MeSH
- DNA nádorová biosyntéza MeSH
- faktor vyvolávající apoptózu metabolismus MeSH
- financování organizované MeSH
- glutathion metabolismus MeSH
- HeLa buňky MeSH
- inhibitor p21 cyklin-dependentní kinasy metabolismus MeSH
- intracelulární signální peptidy a proteiny metabolismus MeSH
- JNK mitogenem aktivované proteinkinasy metabolismus MeSH
- kaspasa 3 metabolismus MeSH
- lidé MeSH
- metaloporfyriny MeSH
- mitochondriální proteiny metabolismus MeSH
- mitogenem aktivované proteinkinasy p38 metabolismus MeSH
- nádorový supresorový protein p53 metabolismus MeSH
- nádory děložního čípku enzymologie patologie MeSH
- oxidační stres účinky léků MeSH
- poškození DNA MeSH
- selen farmakologie MeSH
- tvar organely účinky léků MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- MeSH
- acyklovir farmakologie MeSH
- adenin analogy a deriváty farmakologie MeSH
- antibiotická rezistence MeSH
- bromodeoxyuridin farmakologie MeSH
- mutace MeSH
- organofosforové sloučeniny farmakologie MeSH
- Simplexvirus fyziologie genetika účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- antibiotická rezistence MeSH
- antigeny analýza MeSH
- arboviry MeSH
- bromodeoxyuridin farmakologie MeSH
- cytopatogenní efekt virový MeSH
- erytrocyty imunologie MeSH
- ethylethery farmakologie MeSH
- filtrace MeSH
- hemaglutinační testy MeSH
- husy MeSH
- imunodifuze MeSH
- komplement fixační testy MeSH
- kultivační média MeSH
- kultivační techniky MeSH
- kuřecí embryo MeSH
- myši MeSH
- plíce mikrobiologie MeSH
- ptáci MeSH
- replikace viru MeSH
- RNA virová analýza MeSH
- srdce mikrobiologie MeSH
- testy inhibice hemaglutinace MeSH
- žlučové kyseliny a soli farmakologie MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- myši MeSH
- zvířata MeSH
- MeSH
- antibiotická rezistence MeSH
- arboviry izolace a purifikace patogenita účinky léků MeSH
- bromodeoxyuridin farmakologie MeSH
- cytopatogenní efekt virový MeSH
- erytrocyty imunologie MeSH
- ethylethery farmakologie MeSH
- filtrace MeSH
- hemaglutinační testy MeSH
- husy MeSH
- imunodifuze MeSH
- komplement fixační testy MeSH
- kultivační média MeSH
- kultivační techniky MeSH
- kuřecí embryo MeSH
- myši MeSH
- plíce mikrobiologie MeSH
- protilátky analýza MeSH
- ptáci MeSH
- replikace viru MeSH
- RNA virová analýza MeSH
- srdce mikrobiologie MeSH
- testy inhibice hemaglutinace MeSH
- žlučové kyseliny a soli farmakologie MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- myši MeSH
- zvířata MeSH
