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

Cited by 5 publications

(6 citation statements)

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“…Foci represent the DSBs in a 1:1 manner and are used as a DNA damage biomarker. Other DNA repair markers include RAD51, 53BP1 (p53-binding protein 1), phospho-p53, and PARP1 [170]. However, the specificity whether such DSB markers recognize only DSBs is a controversial issue.…”

mentioning

confidence: 99%

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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mentioning

confidence: 99%

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

Tremi

Spyratou

Souli

et al. 2021

Cancers

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“…Double-strand breaks are a type of DNA damage that can be determined by measuring the number of γH2aX foci [ 33 ]. In the present study, we first explored luteolin-induced DNA damage by measuring the number of γH2aX foci.…”

Section: Resultsmentioning

confidence: 99%

Luteolin impacts deoxyribonucleic acid repair by modulating the mitogen-activated protein kinase pathway in colorectal cancer

Song¹,

Wang³

et al. 2022

Bioengineered

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This study aimed to investigate the effects of luteolin on colorectal cancer (CRC) and explore its underlying mechanism. HCT-116 and HT-29 cells were treated with luteolin, cisplatin, or selumetinib. The cell survival, cell proliferation, apoptosis and cell cycle distribution, and DNA damage were detected using Cell Counting Kit-8, colony formation, flow cytometry, and immunofluorescence staining analysis, respectively. Western blotting was used to detect the expression of apoptosis-related, cycle-related, DNA-damage-related, and mitogen-activated protein kinase (MAPK) pathway-related proteins. Luteolin showed inhibitory effects on cellular growth by reducing cell survival and proliferation, inducing apoptosis and DNA damage, and arresting the cell cycle in a concentration-dependent manner in HCT-116 and HT-29 cells. Meanwhile, luteolin increased the expression of pro-apoptotic proteins, p-CHK1 (central to the induction of cell cycle arrest), and DNA excision repair protein and decreased anti-apoptotic proteins, G2-M phase-related proteins, and DNA repair proteins. The combination of cisplatin and luteolin significantly decreased cell survival and increased the apoptosis rate of HCT-116 and HT-29 cells compared with cisplatin alone. Bioinformatic analysis using the Comparative Toxicogenomics Database and STITCH and MalaCards databases showed that the MAPK pathway is involved in the pharmacology of luteolin. Furthermore, western blotting demonstrated that luteolin plays an inhibitory role by suppressing the MAPK signaling pathway in CRC, which is enhanced when combined with selumetinib. Luteolin can also prevent tumourigenesis in CRC in vivo. In conclusion, luteolin suppressed cell proliferation, blocked the cell cycle, and induced DNA damage and apoptosis progression in CRC cells by mediating the MAPK pathway

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“…Phosphorylated DNA-PKcs involved in this pathway plays a crucial role by binding to the DNA ends, and thus, making a choice between the two pathways. Moreover, DNA-PKcs is only recruited to longer-lived complex DSBs and could play an essential role in repair after the BNCT mixed radiation field ( 26 , 34 ).…”

Section: Discussionmentioning

confidence: 99%

“…γ-H2AX is dephosphorylated when DNA repair is completed; therefore, the DSB marker γ-H2AX is studied extensively through the characterization of foci formation, size, and quantity. Ionizing radiation-induced foci (IRIF) are brighter and larger after high-LET exposure compared with low-LET radiation ( 9 , 18 , 26 ). The formation of γ-H2AX foci leads to the recruitment and accumulation of DNA damage response (DDR) proteins and chromatin-modifying factors, such as 53BP1 (P53 binding protein 1), MDC1 (mediator of DNA damage checkpoint), BRCA1 (Breast Cancer 1 protein), Mre11/Rad50/Nbs1, PARP-1 (poly(ADP-ribose) polymerase 1), and many others, thus forming radiation-induced foci and co-localization with γ-H2AX through direct or indirect binding ( 10 , 11 , 27 ).…”

Section: Dna Damage Response and Repair Pathways After The Mixed Radiation Fieldsmentioning

confidence: 99%

Molecular Mechanisms of Specific Cellular DNA Damage Response and Repair Induced by the Mixed Radiation Field During Boron Neutron Capture Therapy

Maliszewska‐Olejniczak

Kaniowski

Araszkiewicz

et al. 2021

Front. Oncol.

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The impact of a mixed neutron-gamma beam on the activation of DNA damage response (DDR) proteins and non-coding RNAs (ncRNAs) is poorly understood. Ionizing radiation is characterized by its biological effectiveness and is related to linear energy transfer (LET). Neutron-gamma mixed beam used in boron neutron capture therapy (BNCT) can induce another type of DNA damage such as clustered DNA or multiple damaged sites, as indicated for high LET particles, such as alpha particles, carbon ions, and protons. We speculate that after exposure to a mixed radiation field, the repair capacity might reduce, leading to unrepaired complex DNA damage for a long period and may promote genome instability and cell death. This review will focus on the poorly studied impact of neutron-gamma mixed beams with an emphasis on DNA damage and molecular mechanisms of repair. In case of BNCT, it is not clear which repair pathway is involved, and recent experimental work will be presented. Further understanding of BNCT-induced DDR mechanisms may lead to improved therapeutic efficiency against different tumors.

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

Cited by 5 publications

References 0 publications

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

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

Luteolin impacts deoxyribonucleic acid repair by modulating the mitogen-activated protein kinase pathway in colorectal cancer

Molecular Mechanisms of Specific Cellular DNA Damage Response and Repair Induced by the Mixed Radiation Field During Boron Neutron Capture Therapy

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