Immunofluorescence Imaging of DNA Damage and Repair Foci in Human Colon Cancer Cells

Maliszewska‐Olejniczak, Kamila; Dróżdż, Agnieszka; Waluś, Martyna; Dorosz, Michał; Gryziński, M.

doi:10.3791/61399-v

Cited by 3 publications

(1 citation statement)

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“…These data indicated that there was an increased activation of the homologous recombination (HR) pathway. However, CX-5461 did not induce the formation of DNA-PKcs (DNA-dependent protein kinase catalytic subunit) foci [26,27], a marker of the non-homologous end joining (NHEJ) pathway (Fig. 4D).…”

Section: Effects Of Cx-5461 On Different Dna Repair Pathwaysmentioning

confidence: 99%

Molecular events leading to CX-5461-induced DNA damage response in vascular smooth muscle cells

Liu

Pan

Zhang

et al. 2023

Preprint

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Background Our previous studies have shown that the novel selective RNA polymerase I inhibitor CX-5461 suppresses proliferation of vascular smooth muscle cells, mainly via induction of activations of ataxia telangiectasia mutated (ATM)/ATM and Rad3-related (ATR) and p53. Currently, there is no information about what kinds of molecular events mediate such a DNA damage response (DDR) in vascular cells. Methods Primary aortic smooth muscle cells were isolated from normal adult Sprague Dawley rats and cultured in vitro. Immunofluorescence assays were conducted to assess the activation of various DDR pathways. Results We demonstrated that CX-5461-induced DDR was not associated with activations of the nucleotide excision repair, DNA mismatch repair, or the non-homologous end joining pathways, while the homologous recombination pathway was activated. However, the alkaline comet assay did not show massive DNA double strand breaks in CX-5461-treated cells. Instead, CX-5461-triggered DDR appeared to be related to induction of DNA replication stress, which was not attributable to increased formation of G-quadruplex or R-loop structures, but might be explained by increased replication-transcription conflict. CX-5461-induced DDR was not exclusively confined to rDNA within the nucleolar compartment; the extra-nucleolar DDR might represent a distinct secondary response related to the downregulated Rad51 expression in CX-5461-treated cells. Conclusions We suggest that DNA replication stress may be the primary molecular event leading to downstream ATM/ATR and p53 activations in CX-5461-treated vascular smooth muscle cells. Our results provide further insights into the molecular mechanisms underlying the reported beneficial effects of CX-5461 in the cardiovascular system.

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Section: Effects Of Cx-5461 On Different Dna Repair Pathwaysmentioning

confidence: 99%

Molecular events leading to CX-5461-induced DNA damage response in vascular smooth muscle cells

Liu

Pan

Zhang

et al. 2023

Preprint

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Novel insights into the role of ion channels in cellular DNA damage response

Maliszewska-Olejniczak,

Bednarczyk

2024

Mutation Research - Reviews in Mutation Research

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Requirements for Designing an Effective Metallic Nanoparticle (NP)-Boosted Radiation Therapy (RT)

Tremi

Spyratou

Souli

et al. 2021

Cancers

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Many different tumor-targeted strategies are under development worldwide to limit the side effects and improve the effectiveness of cancer therapies. One promising method is to enhance the radiosensitization of the cancer cells while reducing or maintaining the normal tissue complication probability during radiation therapy using metallic nanoparticles (NPs). Radiotherapy with MV photons is more commonly available and applied in cancer clinics than high LET particle radiotherapy, so the addition of high-Z NPs has the potential to further increase the efficacy of photon radiotherapy in terms of NP radiosensitization. Generally, when using X-rays, mainly the inner electron shells are ionized, which creates cascades of both low and high energy Auger electrons. When using high LET particles, mainly the outer shells are ionized, which give electrons with lower energies than when using X-rays. The amount of the produced low energy electrons is higher when exposing NPs to heavy charged particles than when exposing them to X-rays. Since ions traverse the material along tracks, and therefore give rise to a much more inhomogeneous dose distributions than X-rays, there might be a need to introduce a higher number of NPs when using ions compared to when using X-rays to create enough primary and secondary electrons to get the desired dose escalations. This raises the questions of toxicity. This paper provides a review of the fundamental processes controlling the outcome of metallic NP-boosted photon beam and ion beam radiation therapy and presents some experimental procedures to study the biological effects of NPs’ radiosensitization. The overview shows the need for more systematic studies of the behavior of NPs when exposed to different kinds of ionizing radiation before applying metallic-based NPs in clinical practice to improve the effect of IR therapy.

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Immunofluorescence Imaging of DNA Damage and Repair Foci in Human Colon Cancer Cells

Cited by 3 publications

References 0 publications

Molecular events leading to CX-5461-induced DNA damage response in vascular smooth muscle cells

Molecular events leading to CX-5461-induced DNA damage response in vascular smooth muscle cells

Novel insights into the role of ion channels in cellular DNA damage response

Requirements for Designing an Effective Metallic Nanoparticle (NP)-Boosted Radiation Therapy (RT)

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