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| 1 | Notch1 downregulation combined with interleukin-24 inhibits invasion and migration of hepatocellular carcinoma cells显示文摘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. | Bing Han Shi-Hai Liu Wei-Dong Guo Bin Zhang Jian-Ping Wang Yu-Kun Cao Jun Liu | 2015 | World Journal of Gastroenterology2015，21，33： | 8 |
| 2 | 肝癌组织中miR-200和Notch1蛋白表达的相关性显示文摘目的探讨肝癌组织中miR-200表达和Notch1蛋白表达的相关性。方法选择36例肝癌及对照癌旁组织,以荧光实时定量RTPCR检测miR-200在癌旁组织及肝癌的各级临床分期中的表达;Western印迹分析肝癌及癌旁组织中Notch1蛋白表达,并进行相关性分析。结果miR-200在肝癌组织中表达量比在癌旁组织中表达低,在转移癌组织中的表达量比非转移癌组织中低;而Notch1蛋白在癌旁组织、非转移癌组织、转移癌组织中的表达依次增加;Notch1蛋白表达和miR-200表达在肝癌组织中呈负相关(P<0.01)。结论 Notch1是miR-200的标靶,可能是miR-200发挥肿瘤抑制作用的重要靶点。 | 江小梅 彭杰文 | 2015 | 中国老年学杂志2015，35，14： | 1 |
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