文章摘要
张令波 张斌 胡伟平 郭拴龙 胡那日苏 付兆臣 高丽 吕智勇.As2O3干预口腔鳞癌A431 细胞EGFR 表达的研究[J].,2014,14(18):3405-3409
As2O3干预口腔鳞癌A431 细胞EGFR 表达的研究
Effects of Arsenic Trioxide on EGFR Expression of Oral SquamousCell Carcinoma A431 Cell
  
DOI:
中文关键词: 三氧化二砷(As2O3)  口腔鳞状细胞癌(OSCC)  表皮生长因子受体(EGFR)  近红外光学成像(NIR)
英文关键词: Arsenic Trioxide (As2O3)  Oral Squamous Cell Carcinoma (OSCC)  Epidermal Growth Factor Receptor (EGFR)  Near Infrared Imaging (NIR)
基金项目:国家自然基金面上项目(81170960);国家自然基金青年项目(81101086);黑龙江省科技厅攻关项目(GC12C303-2); 哈尔滨医科大学附属第二医院青年启动基金(QN2010-04)
作者单位
张令波 张斌 胡伟平 郭拴龙 胡那日苏 付兆臣 高丽 吕智勇 哈尔滨医科大学附属第二医院口腔科
哈尔滨医科大学硬组织发育和再生研究所
黑龙江省医学科学院
中俄合作哈尔滨医科大学硬组织发育与再生研究所 
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中文摘要:
      目的:研究三氧化二砷(As2O3)对人口腔鳞癌A431细胞生长的抑制作用,探讨其抗肿瘤的机制。方法:合成特异性靶向到肿 瘤细胞表面表皮生长因子受体(EGFR)的近红外荧光分子对比剂EGF-Cy5.5,验证试剂合成的靶向特异性。口腔鳞状细胞癌 A431 细胞系暴露于浓度分别为0 滋M,0.5 滋M,2.5 滋M和5.0 滋M的三氧化二砷溶液中0,24 h,48 h和72 h。共聚焦显微镜、流式 细胞仪及免疫组化证实EGFR的表达水平,上述实验均测量三次,结果取平均值。结果:EGF-Cy5.5 靶向荧光对比剂的标记率为 68%~70 %。对比对照组,越高浓度的三氧化二砷处理的肿瘤细胞其获得的细胞荧光信号强度越小,这与药物浓度越高细胞表面表 达EGFR 的量越少相一致。流式细胞仪显示,在72 小时,作用于细胞的三氧化二砷药物浓度分别为0.5 滋M,2.5 滋M,和5.0 滋M, 其相对应获得的细胞EGFR 表达量分别为57.28± 3.2 %(P<0.05), 29.91± 2.2 %(P<0.01) 和10.73± 2.4 %(P<0.01),明显低于对照 组的细胞EGFR 表达量74.42± 1.8 %,(P <0.05)。结论:本研究应用近红外荧光分子成像的方法体外检测口腔鳞状细胞癌A431 的 EGFR表达水平,实验证明三氧化二砷对其EGFR 具有明显的抑制作用,且抑制作用具有时间- 剂量依赖性。
英文摘要:
      Objective:This study was to investigate the inhibitory effect and mechanism of Arsenic Trioxide (As2O3) on oral squamous cell carcinoma A431 cell.Methods:EGF-Cy5.5 was synthesized according to our previous report and was used as a tumor cellular EGFR specific imaging agent. A431 were exposed to 0 uM, 0.5 uM, 2.5 uM, or 5 uM of As2O3 for 0 h, 24 h, 48 h and 72 h. Confocal microscopy and flow cytometry and cell immunohistochemical staining confirmed EGFR expression. All the studies were measured 3 times and the results were presented as mean.Results:The labeling rate of targeted fluorescent contrast agent EGF-Cy5.5 was 68%to 70 % in vitro , less intense cellular fluorescence signal was observed in cells treated with higher concentration of As2O3 compared with control cells, consistent with the lower levels of EGFR expression in treatment cells. Flow cytometry showed the cellular EGFR expression was 57.28± 3.2 %(P<0.05), 29.91± 2.2 %(P<0.01), and 10.73± 2.4 % (P<0.01) in cells treated with 0.5 uM, 2.5 uM, and 5.0 uMAs2O3, respectively, which were significant lower than the control group (74.42± 1.8 %, P<0.05) at 72 hours.Conclusion:This study assessed in vitro by NIR molecular imaging that the EGFR expression of OSCC A431 cell and demonstrates the inhibition effect of OSCC response to As2O3 treatment in dose-dependent characteristics.
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