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
Objective To investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910...Objective To investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910 cells.Methods Activation of the ERK1/2 and p38 was detected by Western blotting using the antibodies against the total ERK1/2 and p38 mitogen-activated protein kinases (MAPKs) protein and the phosphorylated forms of them. Results Dex could suppress the activation of ERK1/2, while enhance the activation of p38 rapidly and strongly in a dose- and time- dependent manner. Neither effect could be blocked by RU486, the antagonist of glucocorticoid receptor (GR).Conclusion Dex has rapid effects on the activation of ERK1/2 and p38, and these effects are not mediated by GR.展开更多
Objective To investigate N-myc downstream-regulated gene 2(NDRG2) expression in ovarian cancer cells and its potential usefulness as a diagnostic marker and/or target for therapeutic intervention.Methods Human NDRG2 L...Objective To investigate N-myc downstream-regulated gene 2(NDRG2) expression in ovarian cancer cells and its potential usefulness as a diagnostic marker and/or target for therapeutic intervention.Methods Human NDRG2 L/S gene was obtained by revers-transcription polymerase chain reaction(RT-PCR). Sequence analysis confirmed the identity of NDRG2 L/S gene, which was then inserted into a eukaryotic vector p LNCX2, which was in turn transfected into NDRG2 gene-negative HO-8910 cells. Flow cytometry(FCM) and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide(MTT) assay were conducted to determine the proliferation rate of HO-8910 cells. Cisplatin resistance of HO-8910 cells transfected with p LNCX2-NDRG2 L/S was evaluated by FCM. Tumors were generated in female nude mice by subcutaneous injection of HO-8910 cells.Results NDRG2 gene was isolated and its expression vector was successfully constructed. NDRG2 expression positively correlated with the proliferation of HO-8910 cells. NDRG2 L/S promoted tumorigenicity in HO-8910 cells.Conclusion The present study identified a novel function of NDRG2 L/S gene and demonstrated its involvement in the promotion of ovarian cancer cell proliferation and enhancement of cisplatin resistance in HO-8910 cells. Future studies are warranted to determine the relationship between NDRG2 upregulation and ovarian cancer progression.展开更多
Objective： The aim of the study was to investigate the mechanism of gemcitabine （GEM） combination with radiation on the high metastasis human ovarian cancer cell line （HO-8910PM）. Methods： Human ovarian cancer c...Objective： The aim of the study was to investigate the mechanism of gemcitabine （GEM） combination with radiation on the high metastasis human ovarian cancer cell line （HO-8910PM）. Methods： Human ovarian cancer cell line HO- 8910PM was treated with different concentrations of gemcitabine for 24 h, then the cells were counted. In the study of GEM combination with radiation, an efficiency of colony formation was observed; the cell cycle and apoptosis were analyzed by flow cytometry; the experiment of depend on the time and its radio sensitivity were observed by using mitotic index with the cells for each 24, 48, 72 and 96 h after experiment. Results： It suggested that the GEM had an inhibition effect on the human ovarian cancer cell line. The alive cell numbers were decreased by following a height of GEM concentration. When GEM in combina- tion with a radiation, the suppression was significantly increased than that of single GEM therapy. The efficiency of colony formation was significantly lower, under this condition the cell could be arrested at G0-G1 phase and could be decreased to enter into the S phase; the apoptosis percentage could be significantly increased; especially, under the 4 Gy and 6 Gy doses the cell apoptosis was more obvious. GEM combination with radiation had depended on the time to the cells; mitotic index of the calls in combination group was observed significantly lower than that of single GEM therapy or single radiation, and this showed that it had an effect of radiosensitivity. Conclusion： The GEM has a significant growth inhibition on the human ovarian cancer cells, GEM combination with radiation could induce HO-8910PM cell occurred arrested and apoptosis. It has depended on the time and has a radiosensitivity effect. The result shows that it is a better method to treat the human ovarian cancer by using radiotherapy combined with gemcitabine.展开更多
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.
文摘Objective To investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910 cells.Methods Activation of the ERK1/2 and p38 was detected by Western blotting using the antibodies against the total ERK1/2 and p38 mitogen-activated protein kinases (MAPKs) protein and the phosphorylated forms of them. Results Dex could suppress the activation of ERK1/2, while enhance the activation of p38 rapidly and strongly in a dose- and time- dependent manner. Neither effect could be blocked by RU486, the antagonist of glucocorticoid receptor (GR).Conclusion Dex has rapid effects on the activation of ERK1/2 and p38, and these effects are not mediated by GR.
文摘Objective To investigate N-myc downstream-regulated gene 2(NDRG2) expression in ovarian cancer cells and its potential usefulness as a diagnostic marker and/or target for therapeutic intervention.Methods Human NDRG2 L/S gene was obtained by revers-transcription polymerase chain reaction(RT-PCR). Sequence analysis confirmed the identity of NDRG2 L/S gene, which was then inserted into a eukaryotic vector p LNCX2, which was in turn transfected into NDRG2 gene-negative HO-8910 cells. Flow cytometry(FCM) and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide(MTT) assay were conducted to determine the proliferation rate of HO-8910 cells. Cisplatin resistance of HO-8910 cells transfected with p LNCX2-NDRG2 L/S was evaluated by FCM. Tumors were generated in female nude mice by subcutaneous injection of HO-8910 cells.Results NDRG2 gene was isolated and its expression vector was successfully constructed. NDRG2 expression positively correlated with the proliferation of HO-8910 cells. NDRG2 L/S promoted tumorigenicity in HO-8910 cells.Conclusion The present study identified a novel function of NDRG2 L/S gene and demonstrated its involvement in the promotion of ovarian cancer cell proliferation and enhancement of cisplatin resistance in HO-8910 cells. Future studies are warranted to determine the relationship between NDRG2 upregulation and ovarian cancer progression.
文摘Objective： The aim of the study was to investigate the mechanism of gemcitabine （GEM） combination with radiation on the high metastasis human ovarian cancer cell line （HO-8910PM）. Methods： Human ovarian cancer cell line HO- 8910PM was treated with different concentrations of gemcitabine for 24 h, then the cells were counted. In the study of GEM combination with radiation, an efficiency of colony formation was observed; the cell cycle and apoptosis were analyzed by flow cytometry; the experiment of depend on the time and its radio sensitivity were observed by using mitotic index with the cells for each 24, 48, 72 and 96 h after experiment. Results： It suggested that the GEM had an inhibition effect on the human ovarian cancer cell line. The alive cell numbers were decreased by following a height of GEM concentration. When GEM in combina- tion with a radiation, the suppression was significantly increased than that of single GEM therapy. The efficiency of colony formation was significantly lower, under this condition the cell could be arrested at G0-G1 phase and could be decreased to enter into the S phase; the apoptosis percentage could be significantly increased; especially, under the 4 Gy and 6 Gy doses the cell apoptosis was more obvious. GEM combination with radiation had depended on the time to the cells; mitotic index of the calls in combination group was observed significantly lower than that of single GEM therapy or single radiation, and this showed that it had an effect of radiosensitivity. Conclusion： The GEM has a significant growth inhibition on the human ovarian cancer cells, GEM combination with radiation could induce HO-8910PM cell occurred arrested and apoptosis. It has depended on the time and has a radiosensitivity effect. The result shows that it is a better method to treat the human ovarian cancer by using radiotherapy combined with gemcitabine.
