AIM: To confirm the anti-invasion and anti-migration effects of down-regulation of Notch1 combined with interleukin(IL)-24 in hepatocellular carcinoma(HCC) cells.METHODS: γ-secretase inhibitors(GSIs) were used to dow...AIM: To confirm the anti-invasion and anti-migration effects of down-regulation of Notch1 combined with interleukin(IL)-24 in hepatocellular carcinoma(HCC) cells.METHODS: γ-secretase inhibitors(GSIs) were used to down-regulate Notch1.Hep G2 and SMMC7721 cells were seeded in 96-well plates and treated with GSI-I or/and IL-24 for 48 h.Cell viability was measured by MTT assay.The cellular and nuclear morphology was observed under a fluorescence microscope.To further verify the apoptotic phenotype,cell cultures were also analyzed by flow cytometry with Annexin V-FITC/propidium iodide staining.The expression of Notch1,SNAIL1,SNAIL2,E-cadherin,IL-24,XIAP and VEGF was detected by Western blot.The invasion and migration capacities of HCC cells were detected by wound healing assays.Notch1 and Snail were downregulated by RNA interference,and the target proteins were analyzed by Western blot.To investigate the mechanism of apoptosis,we analyzed Hep G2 cells treated with si Notch1 or si CON plus IL-24 or not for 48h by caspase-3/7 activity luminescent assay.RESULTS: GSI-I at a dose of 2.5 μmol/L for 24 h caused a reduction in cell viability of about 38% in Hep G2 cells.The addition of 50 ng/m L IL-24 in combination with 1 or 2.5 μmol/L GSI-I reduced cell viability of about 30% and 15%,respectively.Treatment with IL-24 alone did not induce any cytotoxic effect.In SMMC7721 cells with the addition of IL-24 to GSI-I(2.5 μmol/L),the reduction of cell viability was only about 25%.Following GSI-I/IL-24 combined treatment for 6 h,the apoptotic rate of Hep G2 cells was 47.2%,while no significant effect was observed in cells treated with the compounds employed separately.Decreased expression of Notch1 and its associated proteins SNAIL1 and SNAIL2 was detected in Hep G2 cells.Increased E-cadherin protein expression was noted in the presence of IL-24 and GSI-I.Furthermore,the increased GSI-I and IL-24 in Hep G2 cell was associated with downregulation of MMP-2,XIAP and VEGF.In the absence of treatment,Hep G2 cells could migrate into the scratched space in 24 h.With IL-24 or GSI-I treatment,the wound was still open after 24 h.And the distance of the wound closure strongly correlated with the concentrations of IL-24 and GSI-I.Treatment of Notch-1 silenced Hep G2 cells with 50 ng/m L IL-24 alone for 48 h induced cytotoxic effects very similar to those observed in non-silenced cells treated with GSI-I/IL-24 combination.Caspase-3/7 activity was increased in the presence of si Notch1 plus IL-24 treatment.CONCLUSION: Down-regulation of Notch1 by GSI-I or si RNA combined with IL-24 can sensitize apoptosis and decrease the invasion and migration capabilities of Hep G2 cells.展开更多
AIM:To assess the significance of interleukin(IL)-24 and vascular endothelial growth factor(VEGF)expression in lymph-node-positive rectal cancer. METHODS:Between 1998 and 2005,90 rectal adenocarcinoma patients with ly...AIM:To assess the significance of interleukin(IL)-24 and vascular endothelial growth factor(VEGF)expression in lymph-node-positive rectal cancer. METHODS:Between 1998 and 2005,90 rectal adenocarcinoma patients with lymph node involvement were enrolled.All patients received radical surgery and postoperative pelvic chemoradiotherapy of 50.4-54.0 Gy.Chemotherapy of 5-fluorouracil and leucovorin or levamisole was given intravenously during the first and last week of radiotherapy,and then monthly for about 6 mo.Expression of IL-24 and VEGF was evaluated by immunohistochemical staining of surgical specimens, and their relations with patient characteristics and survival were analyzed.The median follow-up of surviving patients was 73 mo(range:52-122 mo). RESULTS:IL-24 expression was found in 81 out of 90 patients;31 showed weak intensity and 50 showedstrong intensity.VEGF expression was found in 64 out of 90 patients.Negative and weak intensities of IL-24 expression were classified as negative expression for analysis.IL-24 expression was significantly reduced in poorly differentiated tumors in comparison with well or moderately differentiated tumors(P=0.004),N2b to earlier N stages(P=0.016),and stageⅢc to stageⅢ a orⅢb(P=0.028).The number of involved lymph nodes was also significantly reduced in IL-24-positive patients in comparison with IL-24-negative ones. There was no correlation between VEGF expression and patient characteristics.Expression of IL-24 and VEGF was not correlated with survival,but N stage and stages were significantly correlated with survival. CONCLUSION:IL-24 expression was significantly correlated with histological differentiation,and inversely correlated with the degree of lymph node involvement in stageⅢrectal cancer.展开更多
基金Supported by Shandong Provincial Natural Science Foundation,China,No.ZR2012HQ039 and No.ZR2014HP065National Natural Science Foundation of China,No.81373172 and No.81402579
文摘AIM: To confirm the anti-invasion and anti-migration effects of down-regulation of Notch1 combined with interleukin(IL)-24 in hepatocellular carcinoma(HCC) cells.METHODS: γ-secretase inhibitors(GSIs) were used to down-regulate Notch1.Hep G2 and SMMC7721 cells were seeded in 96-well plates and treated with GSI-I or/and IL-24 for 48 h.Cell viability was measured by MTT assay.The cellular and nuclear morphology was observed under a fluorescence microscope.To further verify the apoptotic phenotype,cell cultures were also analyzed by flow cytometry with Annexin V-FITC/propidium iodide staining.The expression of Notch1,SNAIL1,SNAIL2,E-cadherin,IL-24,XIAP and VEGF was detected by Western blot.The invasion and migration capacities of HCC cells were detected by wound healing assays.Notch1 and Snail were downregulated by RNA interference,and the target proteins were analyzed by Western blot.To investigate the mechanism of apoptosis,we analyzed Hep G2 cells treated with si Notch1 or si CON plus IL-24 or not for 48h by caspase-3/7 activity luminescent assay.RESULTS: GSI-I at a dose of 2.5 μmol/L for 24 h caused a reduction in cell viability of about 38% in Hep G2 cells.The addition of 50 ng/m L IL-24 in combination with 1 or 2.5 μmol/L GSI-I reduced cell viability of about 30% and 15%,respectively.Treatment with IL-24 alone did not induce any cytotoxic effect.In SMMC7721 cells with the addition of IL-24 to GSI-I(2.5 μmol/L),the reduction of cell viability was only about 25%.Following GSI-I/IL-24 combined treatment for 6 h,the apoptotic rate of Hep G2 cells was 47.2%,while no significant effect was observed in cells treated with the compounds employed separately.Decreased expression of Notch1 and its associated proteins SNAIL1 and SNAIL2 was detected in Hep G2 cells.Increased E-cadherin protein expression was noted in the presence of IL-24 and GSI-I.Furthermore,the increased GSI-I and IL-24 in Hep G2 cell was associated with downregulation of MMP-2,XIAP and VEGF.In the absence of treatment,Hep G2 cells could migrate into the scratched space in 24 h.With IL-24 or GSI-I treatment,the wound was still open after 24 h.And the distance of the wound closure strongly correlated with the concentrations of IL-24 and GSI-I.Treatment of Notch-1 silenced Hep G2 cells with 50 ng/m L IL-24 alone for 48 h induced cytotoxic effects very similar to those observed in non-silenced cells treated with GSI-I/IL-24 combination.Caspase-3/7 activity was increased in the presence of si Notch1 plus IL-24 treatment.CONCLUSION: Down-regulation of Notch1 by GSI-I or si RNA combined with IL-24 can sensitize apoptosis and decrease the invasion and migration capabilities of Hep G2 cells.
文摘AIM:To assess the significance of interleukin(IL)-24 and vascular endothelial growth factor(VEGF)expression in lymph-node-positive rectal cancer. METHODS:Between 1998 and 2005,90 rectal adenocarcinoma patients with lymph node involvement were enrolled.All patients received radical surgery and postoperative pelvic chemoradiotherapy of 50.4-54.0 Gy.Chemotherapy of 5-fluorouracil and leucovorin or levamisole was given intravenously during the first and last week of radiotherapy,and then monthly for about 6 mo.Expression of IL-24 and VEGF was evaluated by immunohistochemical staining of surgical specimens, and their relations with patient characteristics and survival were analyzed.The median follow-up of surviving patients was 73 mo(range:52-122 mo). RESULTS:IL-24 expression was found in 81 out of 90 patients;31 showed weak intensity and 50 showedstrong intensity.VEGF expression was found in 64 out of 90 patients.Negative and weak intensities of IL-24 expression were classified as negative expression for analysis.IL-24 expression was significantly reduced in poorly differentiated tumors in comparison with well or moderately differentiated tumors(P=0.004),N2b to earlier N stages(P=0.016),and stageⅢc to stageⅢ a orⅢb(P=0.028).The number of involved lymph nodes was also significantly reduced in IL-24-positive patients in comparison with IL-24-negative ones. There was no correlation between VEGF expression and patient characteristics.Expression of IL-24 and VEGF was not correlated with survival,but N stage and stages were significantly correlated with survival. CONCLUSION:IL-24 expression was significantly correlated with histological differentiation,and inversely correlated with the degree of lymph node involvement in stageⅢrectal cancer.
