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Objective@#To explore the effects of FOXO4 D-retro-inverso peptide (FOXO4-DRI) on the radiosensitivity of non-small cell lung cancer (NSCLC) cells.@*Methods@#To detect the effect of FOXO4-DRI on NSCLC cells, H460 and A549 human lung cancer cells were divided into four groups, including untreated control, FOXO4-DRI, γ-ray irradiation and FOXO4-DRI + γ-ray groups. A sigle dose rate of 0.99 Gy γ-rays was used for radiation. H460 cells were administered with 6 μmol/L FOXO4-DRI and A549 cells were adiminstered with 30 μmol/L FOXO4-DRI at 10 min before radiation. Cell viability and survival were detected by CCK-8 assay and colony formation assay, respectively. Cell migration was detected by wound healing assay. Apoptosis and cell cycle arrest were detected with flow cytometry.@*Results@#FOXO4-DRI inhibited growth of H460 and A549 cells (t=1.06-50.75, P<0.05), and decreased cell mobility (t=33.37-139.10, P<0.05) and colony formation (t=5.20-93.48, P<0.05). FOXO4-DRI also increased apoptosis (t=2.95-42.00, P<0.05) and caused a cell cycle arrest at G0/G1 phase accompanied with a decreased proportion of G2/M phase (t=3.50-31.59, P<0.05). Furthermore, FOXO4-DRI increased radiosensitivity of both H460 cells and A549 cells (t=2.94-23.40, P<0.05), caused a Further Decrease of radiation-mediated mobility (t=5.25, 7.56, P<0.05) and colony formation (t=8.43-34.00, P<0.05) and a more increase of radiation-induced apoptosis (t=9.20-11.52, P<0.05). FOXO4-DRI also further decreased the proportion of G2/M phase cells but increased the proportion of S phase cells (t=3.85-17.62, P<0.05).@*Conclusion@#FOXO4-DRI increases radiosensitivity of NSCLC cells by inducing apoptosis and suppressing cell proliferation.
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Objective To explore the effects of FOXO4 D-retro-inverso peptide (FOXO4-DRI)on the radiosensitivity of non-small cell lung cancer (NSCLC) cells.Methods To detect the effect of FOXO4-DRI on NSCLC cells,H460 and A549 human lung cancer cells were divided into four groups,including untreated control,FOXO4-DRI,γ-ray irradiation and FOXO4-DRI + γ-ray groups.A sigle dose rate of 0.99 Gy γ-rays was used for radiation.H460 cells were administered with 6 μmol/L FOXO4-DRI and A549 cells were adiminstered with 30 μmol/L FOXO4-DRI at 10 min before radiation.Cell viability and survival were detected by CCK-8 assay and colony formation assay,respectively.Cell migration was detected by wound healing assay.Apoptosis and cell cycle arrest were detected with flow cytometry.Results FOXO4-DRI inhibited growth of H460 and A549 cells (t =1.06-50.75,P＜ 0.05),and decreased cell mobility (t =33.37-139.10,P＜0.05) and colony formation (t =5.20-93.48,P＜0.05).FOXO4-DRI also increased apoptosis (t=2.95-42.00,P＜0.05) and caused a cell cycle arrest at G0/G1 phase accompanied with a decreased proportion of G2/M phase (t =3.50-3 1.59,P＜0.05).Furthermore,FOXO4-DRI increased radiosensitivity of both H460 cells and A549 cells (t =2.94-23.40,P＜0.05),caused a Further Decrease of radiation-mediated mobility (t =5.25,7.56,P＜0.05) and colony formation (t =8.43-34.00,P＜ 0.05) and a more increase of radiation-induced apoptosis (t =9.20-11.52,P ＜0.05).FOXO4-DRI also further decreased the proportion of G2/M phase cells but increased the proportion of S phase cells (t =3.85-17.62,P ＜ 0.05).Conclusion FOXO4-DRI increases radiosensitivity of NSCLC cells by inducing apoptosis and suppressing cell proliferation.
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Objective To observe the protective effect of anthocyanin on irradiation induced bone marrow c-kit positive cell injury, and further explore its possible mechanism. Methods Mouse bone marrow c-kit positive cells were collected by cell sorting method. There were 2 groups: control group and anthocyanin group, which were sub-divided into three groups and received 0 Gy, 1 Gy and 4 Gy irradiation respectively. The control group was added 700μL cell suspension and an equal volume of serum-free hematopoietic stem/progenitor cell culture medium. The 2 × 10-5 mol/L anthocyanin was co-cultured with mouse bone marrow c-kit positive cells of anthocyanin group half an hour before irradiation exposure, then cells were cultured for 18 hours under the conventional culture conditions (37℃,5%CO2). Mouse c-kit positive cell viability was measured by bioluminescence, and which was reflected by relative light units (RLU). The ability of colony-forming units was reflected by CFU-GM. The reactive oxygen species (ROS) level and mean fluorescence intensity (MFI) ofγ-H2AX were detected by flow cytometry. Results Compared to un-irradiated control group, the cell viability and the number of CFU-GM were decreased significantly, while the ROS level and MFI ofγ-H2AX were increased in c-kit positive cells irradiated with 1 Gy and 4 Gy (P<0.05). Compared to 1 Gy and 4 Gy irradiation groups, c-kit positive cell viability and the number of CFU-GM were increased, the ROS level and MFI of γ-H2AX were decreased in anthocyanin group (P < 0.05). Conclusion Anthocyanin exhibits a promising protective effect on radiation-induced bone marrow c-kit positive cell injury, which may be related to the alleviating ROS and DNA damage in bone marrow cells.
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Objective To investigate the protective effect of theaflavins on thymus injury caused by total body irradiation (TBI).Methods Twenty-five C57BL/6 mice were randomly divided into 5 groups:control group,4 Gy TBI group,4 Gy TBI + 25 mg/kg theaflavins group,4 Gy TBI + 50 mg/kg theaflavins group and 4 Gy TBI + 100 mg/kg theaflavins group.Thymus index and total number of thymocytes were detected at the 14th d post-irradiation to determine the optimal dose of theaflavins.According to this optimal dose,32 C57BL/6 mice were randomly divided into 4 groups:control group,theaflavins group,4 Gy TBI group and 4 Gy TBI + theaflavins group.Thymus histomorphology,CD4CD8 T cell subsets,and reactive oxygen species (ROS) in thymocytes were examined at the 14th d post-irradiation.Results The irradiated thymus exhibited decreased thymus index and total number of thymocytes (P ＜ 0.05),aberrant histomorphology and T cell subsets (P ＜ 0.05),and increased ROS level in thymocytes (P ＜ 0.05).Compared with 4 Gy TBI group,the thymus index and total number of thymocytes were significantly increased in 4 Gy TBI + 50 mg/kg theaflavins group (P ＜ 0.05).The total number of thymocytes was significantly higher in 4 Gy TBI + 50 mg/kg theaflavins group than that in 4 Gy TBI + 25 mg/kg theaflavins group (P ＜ 0.05).Therefore,50 mg/kg theaflavins was chosen as the optimal dose for subsequent experiments.Moreover,the aberrant histomorphology of irradiated thymus was alleviated by theaflavins.A decline in the percentage of CD4-CD8-T cells and an elevation of CD4+CD8-and CD4+CD8+ T cells were found in irradiated mice administered with theaflavins (P ＜ 0.05).Compared with 4 Gy TBI group,the ROS level was significantly decreased in 4 Gy TBI + theaflavins group (P ＜ 0.05).Conclusion Theaflavins exhibits a protective effect on radiation-induced thymus injury.
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Objective To investigate the protective effect of hydrogen-rich water (HRW) on radiation-induced hematopoietic stem and progenitor cells (HSPCs) injury.Methods Totally 32 C57BL/6 mice were randomly divided into four groups with 8 mice in each group,including control,HRW,radiation and radiation + HRW.Mice in HRW and radiation + HRW groups received 0.5 ml hydrogen-rich water per day by intragastric administration 5 min before irradiation until 7 d post-irradiation.Mice in other groups received 0.5 ml distilled water.Mice in radiation and radiation + HRW group were irradiated with 2 Gy of total body irradiation.Bone marrow cells were isolated at 15 d post-irradiation,and LSK cells were examined for the percentage of hematopoietic stem and progenitor cells,the ability of colony formation and reconstitution,reactive oxygen species (ROS) levels and cell apoptosis.Results Compared with radiation group,the percentages of hematopoietic progenitor cells and LSK cells,colony number of bone marrow cells were significantly increased in radiation + HRW group (t =-4.935,-7.898,5.488,P ＜ 0.05).An elevation of donor chimerism was also found in recipient mice administered HRW after competitive bone marrow transplantation (t =-12.769,P ＜ 0.05).Compared with radiation group,the ROS levels and cell apoptosis in LSK cells were significantly decreased (t =4.380,3.954,P ＜ 0.05).Conclusions Hydrogen-rich water exhibited a protective effect on radiation-induced HSPCs injury.
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Objective To evaluate the performance of the Sysmex XE-5000 analyzer for analyzing atypical lymphocytes ,baso-philic granulocytes and their abnormalities warnings .Methods A total of 197 specimens with both atypical lymphocytes and baso-philic granulocytes warnings and 914 specimens with single warning of atypical lymphocytes indicated by Sysmex-5000 blood cell analyzer were collected and inspected by microscope simultaneously .Results Using microscopic examination as a standard ,baso-philic granulocytes within the normal range ,the coincidence rate of samples with both atypical lymphocytes and basophilic granulo-cytes warnings was 64 .9% ,while the coincidence rate of samples with single warning of atypical lymphocytes was 72 .5% .The for-mer was significantly lower than the latter(P<0 .05) .Conclusion When Sysmex XE-5000 indicates atypical lymphocytes and baso-philic granulocytes simultaneously ,there is interference between each other .It should be combined with microscopic examination in order to reduce the probability of missed diagnosis and misdiagnosis .