The magnitude and time-dependence of the apoptotic response of normal and malignant cells subjected to ionizing radiation versus hyperthermia

Vorotnikova, Ekaterina; Ivkov, Robert; Foreman, Allan R.; Tries, Mark A.; Braunhut, Susan J.

doi:10.1080/09553000600876678

Cited by 35 publications

(30 citation statements)

References 35 publications

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“…Cells in 8-chamber Labteks were treated in parallel, and at 24 h the cell free ECM in labteks was fixed with methanol, rinsed with PBS, blocked with normal serum for 30 min and then visualized using indirect immunohistochemistry with rabbit IgG raised to human fibronectin (Zymed Laboratories, San Franciso, CA) visualized using FITC-labeled anti-rabbit antibody conjugates (1:100). The cellfree ECMs were also stained after methanol fixation using 4′6-diamidino-2-phenlindole to detect DNA, as described previously by us (Vorotnikova et al, 2006). The unfixed flask derived cell-free matrix and the cell preparations derived from MRL-B cultures in the T-150 flasks, were normalized for cell numbers and either incubated in a mammalian cell lysis and extraction RIPA buffer consisting of 25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% Sodium deoxycholate and 0.1% SDS from Pierce Products (Thermo Fisher Scientific, Rockford, IL.)…”

Section: Mrl Blastemal Cell Matrix Isolationmentioning

confidence: 99%

Extracellular matrix-derived products modulate endothelial and progenitor cell migration and proliferation in vitro and stimulate regenerative healing in vivo

et al. 2010

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Section: Mrl Blastemal Cell Matrix Isolationmentioning

confidence: 99%

Extracellular matrix-derived products modulate endothelial and progenitor cell migration and proliferation in vitro and stimulate regenerative healing in vivo

et al. 2010

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“…Images were obtained using fluorescence microscopy (Olympus, Center Valley, PA). Mitotic, apoptotic and interphase DNA morphology could be seen by DAPI as previously reported by us [24,25]. The DAPI images were taken using an excitation wavelength filter of 350 nm and the Lava Cell images were taken using an excitation wavelength filter of 534 nm.…”

Section: Methodsmentioning

confidence: 68%

A living cell quartz crystal microbalance biosensor for continuous monitoring of cytotoxic responses of macrophages to single-walled carbon nanotubes

et al. 2011

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BackgroundNumerous engineered nanomaterials (ENMs) exist and new ENMs are being developed. A challenge to nanotoxicology and environmental health and safety is evaluating toxicity of ENMs before they become widely utilized. Cellular assays remain the predominant test platform yet these methods are limited by using discrete time endpoints and reliance on organic dyes, vulnerable to interference from ENMs. Label-free, continuous, rapid response systems with biologically meaningful endpoints are needed. We have developed a device to detect and monitor in real time responses of living cells to ENMs. The device, a living cell quartz crystal microbalance biosensor (QCMB), uses macrophages adherent to a quartz crystal. The communal response of macrophages to treatments is monitored continuously as changes in crystal oscillation frequency (Δf). We report the ability of this QCMB to distinguish benign from toxic exposures and reveal unique kinetic information about cellular responses to varying doses of single-walled carbon nanotubes (SWCNTs).ResultsWe analyzed macrophage responses to additions of Zymosan A, polystyrene beads (PBs) (benign substances) or SWCNT (3-150 μg/ml) in the QCMB over 18 hrs. In parallel, toxicity was monitored over 24/48 hrs using conventional viability assays and histological stains to detect apoptosis. In the QCMB, a stable unchanging oscillation frequency occurred when cells alone, Zymosan A alone, PBs alone or SWCNTs without cells at the highest dose alone were used. With living cells in the QCMB, when Zymosan A, PBs or SWCNTs were added, a significant decrease in frequency occurred from 1-6 hrs. For SWCNTs, this Δf was dose-dependent. From 6-18 hrs, benign substances or low dose SWCNT (3-30 μg/ml) treatments showed a reversal of the decrease of oscillation frequency, returning to or exceeding pre-treatment levels. Cell recovery was confirmed in conventional assays. The lag time to see the Δf reversal in QCMB plots was linearly SWCNT-dose dependent. Lastly, the frequency never reversed at high dose SWCNT (100-150 μg/ml), and apoptosis/necrosis was documented in conventional 24 and 48 hr-assays.ConclusionThese data suggest that the new QCMB detects and provides unique information about peak, sub-lethal and toxic exposures of living cells to ENMs before they are detected using conventional cell assays.

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“…It has been utilized in stereotactic radiotherapy for the treatment of intracranial or body trunk tumors [1], but little is known concerning histological changes after such high-dose irradiation. Laboratory studies have suggested that cell death after high-dose irradiation occurs mainly by apoptosis rather than necrosis [2][3][4]. After irradiating human promyelocytic leukemia cells with 10 or 50 Gy of X rays, Dynlacht et al studied the cells for up to 72 h. After 50 Gy irradiation, cells were found to die primarily by apoptosis, while cells irradiated with 10 Gy died predominantly by necrosis [2].…”

Section: Introductionmentioning

confidence: 97%

“…After irradiating human promyelocytic leukemia cells with 10 or 50 Gy of X rays, Dynlacht et al studied the cells for up to 72 h. After 50 Gy irradiation, cells were found to die primarily by apoptosis, while cells irradiated with 10 Gy died predominantly by necrosis [2]. Human colorectal carcinoma cells irradiated with 20 Gy by Vorotnikova et al [3] showed apoptosis but not necrosis. However, in vivo studies which investigated histological changes after radiation therapy have failed to offer enough evidence that high-dose irradiation causes apoptosis rather than necrosis [5][6][7].…”

Section: Introductionmentioning

confidence: 97%

Histological changes after single high-dose irradiation for squamous cell carcinoma arising from a burn scar

et al. 2009

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Single high-dose irradiation has become a safe and encouraging form of radiation therapy but variations in the histological changes after irradiation have not been fully understood in clinical settings. The aim of this study was to investigate the histological changes of tumor cells after single high-dose, direct irradiation with a case of cutaneous squamous cell carcinoma that developed on a burn scar. The patient received irradiation followed by radical resection of the tumor. Tissues before and after irradiation were stained with Hematoxylin and Eosin (H&E), TdT-mediated dUTP-X nick end labeling (TUNEL), and by immunohistochemistry for MIB-1, p53, and p21. Massive necrosis was noted after irradiation but a few viable tumor cells remained. Preradiation histology revealed MIB-1 and p53 as positive, and p21 tumor cells as partially positive. These expressions were reduced after irradiation. The TUNEL stain was consistently negative. It was established that tumor cell death after single high-dose irradiation was caused mainly by necrosis rather than apoptosis in clinical settings.

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The magnitude and time-dependence of the apoptotic response of normal and malignant cells subjected to ionizing radiation versus hyperthermia

Abstract: These studies reveal time and temperature dependent in vitro cell responses to ionizing radiation and water-bath hyperthermia.

Cited by 35 publications

References 35 publications

Extracellular matrix-derived products modulate endothelial and progenitor cell migration and proliferation in vitro and stimulate regenerative healing in vivo

Extracellular matrix-derived products modulate endothelial and progenitor cell migration and proliferation in vitro and stimulate regenerative healing in vivo

A living cell quartz crystal microbalance biosensor for continuous monitoring of cytotoxic responses of macrophages to single-walled carbon nanotubes

Histological changes after single high-dose irradiation for squamous cell carcinoma arising from a burn scar

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