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γ-干扰素对喉癌细胞株碱性成纤维细胞生长因子表达的影响

Effect of interferon-gamma on the expression of basic fibroblast growth factor on Hep-2 laryngeal carcinoma cell lines
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摘要 目的:研究γ-干扰素(IFN-γ)对人喉癌细胞株Hep-2细胞碱性成纤维细胞生长因子(bFGF)基因及蛋白表达的影响。方法:将IFN-γ以不同浓度[(1、10、100、1000、10000)U/ml]、不同作用时间[(0、12、24、36、48、60、72)h]作用于Hep-2细胞,用四甲基偶氮唑蓝(MTT)比色法测定细胞增殖;用实时荧光定量PCR(Realtime-PCR)法测定IFN-γ(1000U/ml)不同作用时间[(0、2、6、12、24、36、48、60、72)h]细胞bFGFmRNA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与Realtime-PCR相同药物剂量和作用时间细胞培养上清液和细胞浆中bFGF蛋白含量。结果:细胞生长受到抑制,以高剂量组表现最为明显。不同浓度(1、10、100、1000、10000U/ml)IFN-γ作用不同时间[(0、12、24、36、48、60、72)h]后,从48h开始,(100、1000、10000)U/ml的IFN-γ对Hep-2细胞有明显的抑制作用,差异有显著性(P<0.05),但不随着浓度的增加而成正比。1000U/mlIFN-γ作用后,在2h后与对照组比,bFGFmRNA表达上调,差异有显著性(P<0.05),特别是48h后,有显著性差异(P<0.01)。1000U/mlIFN-γ作用48h后,与对照组比较,Hep-2细胞浆中的棕色颗粒明显增多。结论:IFN-γ抑制Hep-2细胞分泌bFGF却,不抑制其合成。 Objective：To observe the effect of interferon-gamma on the expression of basic fibroblast growth factor on Hep-2 laryngeal carcinoma cell lines.Methods：Hep-2 cell was exposed to interferon-gamma at a concentration（1,10,100,1 000,10 000）U/ml for（0,12,24,36,48,60,72）h MTT,colorimetric assay was used to eva1uate the effect of interferon-gamma on Hep-2 cell proliferation after incubation with interferon-gamma（1,10,100,1000,10 000）U/ml for（0,12,24,36,48,60,72）h.Hep-2 cell was harvested to detect bFGF mRNA levels by realtime polymerase chain reaction（realtime-PCR） with interferon-gamma（1000U/ml）for（0,2,6,12,24,36,48,60,72）h.The bFGF protein level in supernatants was determined by enzyme-l inked immunosorbent assay（ELISA） and in cytoplasma by immunocytochemical staining with the same drug treatment as realtime-PCR.Results：Hep-2 cell was significantly inhibited by IFN-γ at high concentration.Expression of bFGF mRNA and protein levels of Hep-2 cell at 1000U/ml concentratlons of interferon-gamma were higher than that of normal control（P〈0.05） at 48h.Conclusion：IFN-γ can inhibit the expression of bFGF in Hep-2 cell but not inhibit its synthesis.
作者 陈婕 周佳青 王家东
机构地区 上海交通大学医学院附属仁济医院耳鼻咽喉-头颈外科
出处 《现代肿瘤医学》 CAS 2007年第10期1385-1388,共4页 Journal of Modern Oncology
关键词 Γ-干扰素 喉癌 碱性成纤维生长因子 interferon-gamma laryngeal carcinomas basic fibroblast growth factor（bFGF）
分类号 R730 [医药卫生—肿瘤]
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参考文献10

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现代肿瘤医学
2007年 第10期

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