Radiation Effects and Radioprotection in MC3T3-E1 Mouse Calvarial Osteoblastic Cells

Gevorgyan, Artur; Sukhu, Balram; Alman, Benjamin A.; Bristow, Robert G.; Pang, Cho Y.; Forrest, Christopher R.

doi:10.1097/prs.0b013e3181845931

Cited by 16 publications

(16 citation statements)

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“…1C.). However, another study analyzed the effect of radiation on OPN levels in osteoblastic cells and found a slightly elevated expression of OPN on days 14 and 21 after irradiation [40]. …”

Section: Discussionmentioning

confidence: 99%

Effects of osteopontin inhibition on radiosensitivityof MDA-MB-231 breast cancer cells

et al. 2010

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BackgroundOsteopontin (OPN) is a secreted glycophosphoprotein that is overexpressed in various tumors, and high levels of OPN have been associated with poor prognosis of cancer patients. In patients with head and neck cancer, high OPN plasma levels have been associated with poor prognosis following radiotherapy. Since little is known about the relationship between OPN expression and radiosensitivity, we investigated the cellular and radiation induced effects of OPN siRNA in human MDA-MB-231 breast cancer cells.MethodsMDA-MB-231 cells were transfected with OPN-specific siRNAs and irradiated after 24 h. To verify the OPN knockdown, we measured the OPN mRNA and protein levels using qRT-PCR and Western blot analysis. Furthermore, the functional effects of OPN siRNAs were studied by assays to assess clonogenic survival, migration and induction of apoptosis.ResultsTreatment of MDA-MB-231 cells with OPN siRNAs resulted in an 80% decrease in the OPN mRNA level and in a decrease in extracellular OPN protein level. Transfection reduced clonogenic survival to 42% (p = 0.008), decreased the migration rate to 60% (p = 0.15) and increased apoptosis from 0.3% to 1.7% (p = 0.04). Combination of OPN siRNA and irradiation at 2 Gy resulted in a further reduction of clonogenic survival to 27% (p < 0.001), decreased the migration rate to 40% (p = 0.03) and increased apoptosis to 4% (p < 0.005). Furthermore, OPN knockdown caused a weak radiosensitization with an enhancement factor of 1.5 at 6 Gy (p = 0.09) and a dose modifying factor (DMF10) of 1.1.ConclusionOur results suggest that an OPN knockdown improves radiobiological effects in MDA-MB-231 cells. Therefore, OPN seems to be an attractive target to improve the effectiveness of radiotherapy.

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Section: Discussionmentioning

confidence: 99%

Effects of osteopontin inhibition on radiosensitivityof MDA-MB-231 breast cancer cells

et al. 2010

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“…For SOD1 and GPx amplification [80], PCR conditions were 94°C for 2 minutes followed by 25 cycles of 94°C for 15 seconds, 57°C for 30 seconds, 72°C for 45 seconds annealing, and a final extension of 72°C for 10 minutes. Using methods described by Gevorgyan et al [81] OCN (bone Gla protein) was amplified using 94°C for 2 minutes followed by 25 cycles of 94°C for 1 minute, 58°C for 45 seconds, 72°C for I minute annealing, and a final extension of 72°C for 10 minutes. Similarly, secreted phosphoprotein 1(OPN) was amplified using 94°C for 2 minutes followed by 18 cycles of 95°C for 1 minute, 55°C for 2 minutes, 72°C for I minute annealing, and a final extension of 72°C for 10 minutes [81].…”

Section: Methodsmentioning

confidence: 99%

“…Using methods described by Gevorgyan et al [81] OCN (bone Gla protein) was amplified using 94°C for 2 minutes followed by 25 cycles of 94°C for 1 minute, 58°C for 45 seconds, 72°C for I minute annealing, and a final extension of 72°C for 10 minutes. Similarly, secreted phosphoprotein 1(OPN) was amplified using 94°C for 2 minutes followed by 18 cycles of 95°C for 1 minute, 55°C for 2 minutes, 72°C for I minute annealing, and a final extension of 72°C for 10 minutes [81]. The PCR products were electrophoresed in 2% agarose gels.…”

Section: Methodsmentioning

confidence: 99%

Doxorubicin-Mediated Bone Loss in Breast Cancer Bone Metastases Is Driven by an Interplay between Oxidative Stress and Induction of TGFβ

et al. 2013

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Breast cancer patients, who are already at increased risk of developing bone metastases and osteolytic bone damage, are often treated with doxorubicin. Unfortunately, doxorubicin has been reported to induce damage to bone. Moreover, we have previously reported that doxorubicin treatment increases circulating levels of TGFβ in murine pre-clinical models. TGFβ has been implicated in promoting osteolytic bone damage, a consequence of increased osteoclast-mediated resorption and suppression of osteoblast differentiation. Therefore, we hypothesized that in a preclinical breast cancer bone metastasis model, administration of doxorubicin would accelerate bone loss in a TGFβ-mediated manner. Administration of doxorubicin to 4T1 tumor-bearing mice produced an eightfold increase in osteolytic lesion areas compared untreated tumor-bearing mice (P = 0.002) and an almost 50% decrease in trabecular bone volume expressed in BV/TV (P = 0.0005), both of which were rescued by anti-TGFβ antibody (1D11). Doxorubicin, which is a known inducer of oxidative stress, decreased osteoblast survival and differentiation, which was rescued by N-acetyl cysteine (NAC). Furthermore, doxorubicin treatment decreased Cu-ZnSOD (SOD1) expression and enzyme activity in vitro, and treatment with anti-TGFβ antibody was able to rescue both. In conclusion, a combination therapy using doxorubicin and anti-TGFβ antibody might be beneficial for preventing therapy-related bone loss in cancer patients.

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“…Their study revealed AMF radioprotection with significant improvement of osteoblast-like cell survival. 31 Yurut-Caloglu et al studied the left femora of Wistar albino rats, which they irradiated at a single dose of 20 Gy following AMF pre-treatment. Their histopathological analysis of bone and cartilage revealed a significant reduction in radiation-induced bone and epiphysial cartilage damage in the AMF pre-treated specimens compared to irradiated specimens alone.…”

Section: Discussionmentioning

confidence: 99%

Quantitative Histologic Evidence of Amifostine-Induced Cytoprotection in an Irradiated Murine Model of Mandibular Distraction Osteogenesis

Tchanque-Fossuo

Donneys

Razdolsky

et al. 2012

Plastic and Reconstructive Surgery

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Background Head and neck cancer (HNC) management requires adjuvant radiation therapy (XRT). The authors have previously demonstrated the damaging effect of a human equivalent dose of radiation (HEDR) on a murine mandibular model of distraction osteogenesis (DO). Utilizing quantitative histomorphometry (QHM), our specific aim is to objectively measure the radio-protective effects of Amifostine (AMF) on the cellular integrity and tissue quality of an irradiated and distracted regenerate. Methods Sprague Dawley rats were randomly assigned into 2 groups: XRT/DO and AMF/XRT/DO, which received AMF prior to XRT. Both groups were given HEDR in 5 fractionated doses and underwent a left mandibular osteotomy with bilateral fixator placement. Distraction to 5.1mm was followed by a 28-day consolidation period. Left hemimandibles were harvested. QHM was performed for osteocyte count (Oc), empty lacunae (EL), Bone Volume/Tissue Volume (BV/TV) and Osteoid Volume/Tissue Volume (OV/TV) ratios. Results AMF/XRT/DO exhibited bony bridging as opposed to XRT/DO fibrous unions. QHM analysis revealed statistically significant higher Oc and BV/TV ratio in AMF-treated mandibles compared with irradiated mandibles. There was a corresponding decrease in EL and the ratio of OV/TV between AMF/XRT/DO and XRT/DO. Conclusion We have successfully established the significant osseous cytoprotective and histoprotective capacity of AMF on DO in the face of XRT. AMF-sparing effect on bone cellularity correlated with an increase in bony union and elimination of fibrous union. We posit that the demonstration of similar efficacy of AMF in the clinic may allow the successful implementation of DO as a viable reconstructive option for HNC in the future.

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Radiation Effects and Radioprotection in MC3T3-E1 Mouse Calvarial Osteoblastic Cells

Cited by 16 publications

References 45 publications

Effects of osteopontin inhibition on radiosensitivityof MDA-MB-231 breast cancer cells

Effects of osteopontin inhibition on radiosensitivityof MDA-MB-231 breast cancer cells

Doxorubicin-Mediated Bone Loss in Breast Cancer Bone Metastases Is Driven by an Interplay between Oxidative Stress and Induction of TGFβ

Quantitative Histologic Evidence of Amifostine-Induced Cytoprotection in an Irradiated Murine Model of Mandibular Distraction Osteogenesis

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