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目的 研究γ 干扰素 (IFN γ)对人卵巢癌细胞株SKOV3细胞血管内皮生长因子 (VEGF)表达的影响。方法 将IFN γ以不同浓度 (1、10、10 0、10 0 0、10 0 0 0U/ml)、不同作用时间 (12、2 4、3 6、48、60h)作用于SKOV3细胞 ,在相差显微镜下观察细胞形态的变化 ;用四甲基偶氮唑蓝 (MTT)比色法测定细胞增殖 ;用逆转录聚合酶链反应 (RT PCR)半定量法测定细胞VEGFmRNA含量 ;用双抗体夹心酶联免疫吸附试验 (ELISA)和免疫细胞化学方法测定细胞培养上清液和细胞浆中VEGF蛋白含量。结果 (1) 10 0 0U/mlIFN γ作用后SKOV3细胞形态无明显变化。 (2 )不同浓度 (1、10、10 0、10 0 0、10 0 0 0U/ml)IFN γ作用不同时间 (12、2 4、3 6、48、60h)后 ,SKOV3细胞增殖活性比较 ,差异无显著性 (P>0 0 5 )。 (3 )IFN γ作用后 ,SKOV3细胞VEGFmRNA和蛋白含量明显减少 (P <0 0 1)。IFN γ浓度达到 10U/ml后起作用 [抑制率为 (5 2± 0 6) % ],其后随着浓度的增高 ,IFN γ对VEGFmRNA的抑制作用增强 ,在浓度达到 10 0 0U/ml时 ,抑制作用达到高峰 [抑制率为 (2 1 6± 2 6) % ]。但IFN γ的抑制作用并不存在时间依赖关系 ,用药 2 4h后出现抑制作用 ,并立即达到高峰 [抑制率为 (2 2 4± 1 7) % ],其后抑制作用不随时间的延长而
Objective To study the effects of IFNγ on the expression of vascular endothelial growth factor (VEGF) in human ovarian cancer cell line SKOV3. Methods IFNγ was administered to SKOV3 cells at different concentrations (1,10,10 0,10 0,10 0 0 0 0U / ml) and different action time (12,24,3 6,48,60h) Cell morphology was observed under microscope. Cell proliferation was measured by MTT assay. VEGF mRNA level was determined by reverse transcription-polymerase chain reaction (RT-PCR) semi-quantitative method. The contents of VEGF protein in cell culture supernatant and cytoplasm were determined by ELISA and immunocytochemistry. Results (1) The morphology of SKOV3 cells did not change significantly after treated with 10 0 U / ml IFNγ. (2) The proliferative activity of SKOV3 cells after different concentrations of IFNγ (1, 10, 10, 10, 10, 100 μU / ml) for different times (12,24,36,48,60h) There was no significant difference (P> 0.05). (3) After IFN γ treatment, the content of VEGF mRNA and protein in SKOV3 cells decreased significantly (P <0.01). IFNγ concentration up to 10U / ml play a role [inhibition rate (5 2 ± 0 6)%], then as the concentration increases, IFNγ on VEGFmRNA inhibition enhanced at a concentration of 10 0U / ml , The inhibition peaked [inhibition rate (2 1 6 ± 2 6)%]. However, the inhibitory effect of IFNγ was not time-dependent. After 24 h, the inhibitory effect was observed and reached the peak immediately (the inhibition rate was (22.4 ± 17)%], and the inhibitory effect was not prolonged with time