粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective To investigate if granulocyte-macrophage colony stimulating factor (GM-CSF) gene-modified dendritic cells ( DC) enhance antitumor immunity in vitro. Methods Mice were injected with chemokine ligand 3 (CCL3) via the tail vein. Fresh B220-CD11c+ cells were sorted from the peripheral blood mononuclear cells (PBMCs) and cultured into DCs by cytokines. DCs were transfected with AdGM-CSF gene at different ratios of multiplicity of infection ( MOI) to determine the optimal gene transfection conditions, and the expression of GM-CSF was detected after transfection. The variation of GM-CSF gene-modifiedDCs were analyzed by morphological examination, phenotype analysis, and mixed lymphocyte reaction (MLR). DCs were loaded with gastric cancer antigen obtained by freezing and thawing method. The killing effect of DCs vaccine-stimulated T lymphocytes on gastric cancer cells was assessed by MTT assay. INF-γ production was determined with the INF-γ ELISA kit. Results B220- CD11c+ cells increased obviously after CCL3 injection. The ELISA results showed that after GM-CSF gene modification, DCs could produce high level of GM-CSF. When DCs were transfected with AdGM-CSF gene at MOI equal to 100, the GM-CSF level in culture supematants reached saturation [(130.00±12.61) pg/ml]. After GM-CSF gene-modification, DCs tend to be more maturated as detected by morphological observation and phenotype analysis. At the same time, the capacity of activating the proliferation of allogeneic T lymphocytes was enhanced greatly. T lymphocytes stimulated by DCs transfected with GM-CSF gene showed a specific killing effect on gastric carcinoma cells and produced high level of INF-γ[ ( 1245. 00±13. 75) pg/ml].Conclusion After GM-CSF gene modification, DCs can produce high level of GM-CSF, which tend to be more maturated, and the capacity of activating the proliferation of allogeneic T lymphocytes is enhanced greatly. GM-CSF gene modified DCs can induce specific CTL to target tumor cells in vitro.     

粒细胞-巨噬细胞集落刺激因子基因修饰的树突状细胞疫苗增强体外抗肿瘤免疫效应

Objective: The aim of the study was to investigate whether dendritic cell (DC) precursors, recruited by injection of chemokine ligand 3 (CCL3), induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor (GM-CSF) transfection in mice ex vivo. Methods: The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c+ cells were cultured with cytokines. For adenoviral (Ad)-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of mu...     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.     

转染血管内皮生长因子C反义寡核苷酸对子宫颈癌Hela细胞增生和凋亡的影响

Objective To explore effect on the proliferation and apoptosis of Hela cells in vitro by using vascular endothelial growth factor-C(VEGF-C) antisense oligonucleotides (ASODN). Methods VEGF-C ASODN was transfected into Hela cells by liposome-mediated; the cells transfected with the oligodeoxynuclecotide (SODN) and saline were used as control groups. The efficiency of transfection was detected by fluorescence microscope. The inhibitive rate of cell growth was detected with MTT methods. Apoptosis and cell cycle were evaluated using FCM. The mRNA expression of VEGF-C was determined by RT-PCR, and the expression of VEGF-C was determined by western blotting. Results VEGF-C ASODN had been transfected into Hela cells by liposome-mediated. The index of apoptosis after transfection 48 h was (19.39±1.81)%. G2/M phase cell increased after transfection, meanwhile the expression of VEGF-C degraded on the level of mRNA and protein (P<0.05). Conclusion Transfeeted with VEGF-C ASODN could down-regulate the expression of VEGF on the level of mRNA and protein, block the cell circle, inhibit cell proliferation and induce apoptosis in Hela cells in vitro.