目的:研究辛伐他汀(SIM)对人高转移卵巢癌HO-8910PM细胞Ras信号通路的影响。方法:不同浓度SIM分别作用HO-8910PM细胞48 h后,采用RT-PCR分别检测Ras、HIF-1α、NF-κB(P65)、Rho A和VEGF m RNA的表达,Western blot分别检测Ras、ERK2、-κ...目的:研究辛伐他汀(SIM)对人高转移卵巢癌HO-8910PM细胞Ras信号通路的影响。方法:不同浓度SIM分别作用HO-8910PM细胞48 h后,采用RT-PCR分别检测Ras、HIF-1α、NF-κB(P65)、Rho A和VEGF m RNA的表达,Western blot分别检测Ras、ERK2、-κBα、NF-κB(p65)、HIF-1α和VEGF的蛋白水平,γ-^(32)P掺入法检测MAPK活性。结果 :SIM能使NF-κB(p65)、VEGF和RHo A m RNA表达降低,影响Ras蛋白在胞浆和胞膜间的分布,下调ERK2、HIF-1α、VEGF蛋白表达,并抑制活化的NF-κB往核内易位。结论:SIM抗卵巢癌机制与抑制Ras/MAPK信号传导及其与NF-κB通路间的交叉通讯密切相关。展开更多
目的:探讨藏红花素对卵巢癌HO-8910细胞自噬的影响及其分子机制。方法:用Western印迹法测定内源性LC3B-II蛋白的稳态水平以及MTOR及其下游底物的磷酸化水平。用荧光和共聚焦显微镜检测GFP-LC3B斑点的分布。结果:与对照细胞相比,用不同...目的:探讨藏红花素对卵巢癌HO-8910细胞自噬的影响及其分子机制。方法:用Western印迹法测定内源性LC3B-II蛋白的稳态水平以及MTOR及其下游底物的磷酸化水平。用荧光和共聚焦显微镜检测GFP-LC3B斑点的分布。结果:与对照细胞相比,用不同浓度的藏红花素处理的HO-8910细胞中内源性LC3B-II蛋白的稳态水平和GFP-LC3B斑点的分布以剂量依赖的方式增强。用藏红花素处理HO-8910细胞后,MTOR及其下游底物的磷酸化水平显著降低。结论:藏红花素通过抑制MTOR信号通路促进卵巢癌HO-8910细胞自噬体的形成。Aims: To investigate the mechanism through which crocin influences the autophagy of ovarian cancer HO-8910 cells. Methods: Western blotting assay was used to determine the steady-state levels of endogenous LC3B-II protein and the phosphorylation level of MTOR and its downstream substrates. Fluorescence and confocal microscopy was used to detect the distribution of GFP-LC3B puncta. Results: Compared to the control cells, the steady-state levels of endogenous LC3B-II protein and the distribution of GFP-LC3B puncta were enhanced in the HO-8910 cells treated with various concentration of crocin in a dose-dependent manner. Following treatment of HO-8910 cells with crocin, the phosphorylation level of MTOR and its downstream substrates decreased significantly. Conclusions: Crocin promotes the formation of autophagosome in ovarian cancer HO-8910 cells by inhibiting the MTOR signaling pathway.展开更多
文摘目的:研究辛伐他汀(SIM)对人高转移卵巢癌HO-8910PM细胞Ras信号通路的影响。方法:不同浓度SIM分别作用HO-8910PM细胞48 h后,采用RT-PCR分别检测Ras、HIF-1α、NF-κB(P65)、Rho A和VEGF m RNA的表达,Western blot分别检测Ras、ERK2、-κBα、NF-κB(p65)、HIF-1α和VEGF的蛋白水平,γ-^(32)P掺入法检测MAPK活性。结果 :SIM能使NF-κB(p65)、VEGF和RHo A m RNA表达降低,影响Ras蛋白在胞浆和胞膜间的分布,下调ERK2、HIF-1α、VEGF蛋白表达,并抑制活化的NF-κB往核内易位。结论:SIM抗卵巢癌机制与抑制Ras/MAPK信号传导及其与NF-κB通路间的交叉通讯密切相关。
文摘目的:探讨藏红花素对卵巢癌HO-8910细胞自噬的影响及其分子机制。方法:用Western印迹法测定内源性LC3B-II蛋白的稳态水平以及MTOR及其下游底物的磷酸化水平。用荧光和共聚焦显微镜检测GFP-LC3B斑点的分布。结果:与对照细胞相比,用不同浓度的藏红花素处理的HO-8910细胞中内源性LC3B-II蛋白的稳态水平和GFP-LC3B斑点的分布以剂量依赖的方式增强。用藏红花素处理HO-8910细胞后,MTOR及其下游底物的磷酸化水平显著降低。结论:藏红花素通过抑制MTOR信号通路促进卵巢癌HO-8910细胞自噬体的形成。Aims: To investigate the mechanism through which crocin influences the autophagy of ovarian cancer HO-8910 cells. Methods: Western blotting assay was used to determine the steady-state levels of endogenous LC3B-II protein and the phosphorylation level of MTOR and its downstream substrates. Fluorescence and confocal microscopy was used to detect the distribution of GFP-LC3B puncta. Results: Compared to the control cells, the steady-state levels of endogenous LC3B-II protein and the distribution of GFP-LC3B puncta were enhanced in the HO-8910 cells treated with various concentration of crocin in a dose-dependent manner. Following treatment of HO-8910 cells with crocin, the phosphorylation level of MTOR and its downstream substrates decreased significantly. Conclusions: Crocin promotes the formation of autophagosome in ovarian cancer HO-8910 cells by inhibiting the MTOR signaling pathway.
