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1 mg/ml 5-bromo-4-chloro-3-inolyl-β-d-galactoside anti-cd44 apc-conjugated antibody anti-cd24 pe-conjugated antibody human colon-cancer-initiating cells pheno-red free dmem-f12 cd24-/low/cd44high cell population article download pdf confluent mcf-7-zsgreen-codc monolayer mcf-7 cd24-/low/cd44high cells cd24-/low/cd44high cics markers related subjects x-gal-positive/senescent cells fluorescence-activated cell-sorting nih-gov/ij/] darzynkiewicz cell-type specific behaviour salt-free staining vehicle cellular oncology cd24-/low/cd44high cells cell line continued primary sphere-forming capacity cd24-/low/cd44high population exhibit multi-lineage potency cancer stem cell β-galactosidase-positive cells including breast cancer zsgreen-codc+ cells constituted mcf-7 cell line breast cancer cell heterogeneous monolayer population sphere-forming capacity assays zsgreen-codc-positive cells fl-1h channel exceeded t47d secondary sphere-formation cell stem cell cancer stem cells performed sphere-forming assays zsgreen-codc+ cells increased low proteasome activity cics zsgreen-codc+ showed natl cancer inst breast cancer cells cancer initiating cells stem cell properties breast cancer res measure zsgreen-codc expression 20 mg/ml stock indicating radiation-induced mobilization tumorigenic zsgreen-codc- cells cd24-/low/cd44high β-galactosidase activity

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         headline:Survival and self-renewing capacity of breast cancer initiating cells during fractionated radiation treatment
         description:Recent data indicate a hierarchical organization of many solid cancers, including breast cancer, with a small number of cancer initiating cells (CICs) that have the ability to self-renew and exhibit multi-lineage potency. We, and others, have demonstrated that CICs in breast cancer and glioma are relatively resistant to ionizing radiation if compared to their non-tumorigenic counterparts. However, the extent of the remaining self-renewing capacity of CICs after fractions of radiation is currently unknown. We hypothesized that CICs, in contrast to their non-tumorigenic counterparts, not only survive fractions of ionizing radiation but also retain the CIC phenotype as defined by operational means. We used two marker systems to identify breast CICs (CD24-/low/CD44high, or lack of proteasome activity) and performed sphere-forming assays after multiple clinical fractions of radiation. Lineage tracking was performed by membrane staining. Cell cycle distribution and RNA content were assessed by flow cytometry and senescence was assessed via β-galactosidase staining. We demonstrated that irradiated CICs survived and retained their self-renewal capacity for at least four generations. We show that fractionated radiation not only spared CICs but also mobilized them from a quiescent/G0 phase of the cell cycle into actively cycling cells, while the surviving non-tumorigenic cells were driven into senescence. The breast CIC population retains increased self-renewal capacity over several generations and therefore, we conclude that increases in the number of CICs after sublethal doses of radiation have potential clinical importance. Prevention of this process may lead to improved clinical outcome.
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            Proteasome Activity
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            Cancer Research
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      headline:Survival and self-renewing capacity of breast cancer initiating cells during fractionated radiation treatment
      description:Recent data indicate a hierarchical organization of many solid cancers, including breast cancer, with a small number of cancer initiating cells (CICs) that have the ability to self-renew and exhibit multi-lineage potency. We, and others, have demonstrated that CICs in breast cancer and glioma are relatively resistant to ionizing radiation if compared to their non-tumorigenic counterparts. However, the extent of the remaining self-renewing capacity of CICs after fractions of radiation is currently unknown. We hypothesized that CICs, in contrast to their non-tumorigenic counterparts, not only survive fractions of ionizing radiation but also retain the CIC phenotype as defined by operational means. We used two marker systems to identify breast CICs (CD24-/low/CD44high, or lack of proteasome activity) and performed sphere-forming assays after multiple clinical fractions of radiation. Lineage tracking was performed by membrane staining. Cell cycle distribution and RNA content were assessed by flow cytometry and senescence was assessed via β-galactosidase staining. We demonstrated that irradiated CICs survived and retained their self-renewal capacity for at least four generations. We show that fractionated radiation not only spared CICs but also mobilized them from a quiescent/G0 phase of the cell cycle into actively cycling cells, while the surviving non-tumorigenic cells were driven into senescence. The breast CIC population retains increased self-renewal capacity over several generations and therefore, we conclude that increases in the number of CICs after sublethal doses of radiation have potential clinical importance. Prevention of this process may lead to improved clinical outcome.
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         T47D Cell
         Proteasome Activity
         Breast Cancer Stem Cell
         T47D Cell Line
         T47D Breast Cancer Cell
         Cancer Research
         Oncology
         Surgical Oncology
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      name:Division of Molecular and Cellular Oncology, Department of Radiation Oncology, David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, USA
      name:Division of Molecular and Cellular Oncology, Department of Radiation Oncology, David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, USA
      name:Division of Molecular and Cellular Oncology, Department of Radiation Oncology, David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, USA
      name:Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, USA

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