| 李建成,张和平,刘迪,陈诚,苏颖,王玲华.表皮生长因子受体siRNA对食管鳞癌细胞的放射增敏研究[J].中华放射医学与防护杂志,2013,33(1):23-26 |
| 表皮生长因子受体siRNA对食管鳞癌细胞的放射增敏研究 |
| Study of siRNA EGFR on the enhanced radiosensitivity of esophageal squamous cells |
| 投稿时间：2012-07-09 |
| DOI：10.3760/cma.j.issn.0254-5098.2013.01.006 |
| 中文关键词: RNA干扰 放射敏感性 表皮生长因子受体 食管癌 |
| 英文关键词:RNA interfering Radiosensitivity EGFR Esophageal neoplasm |
| 基金项目:福建省卫生创新课题(2007-CX-4) |
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| 中文摘要: |
| 目的 探讨用小干扰RNA(siRNA)抑制表皮生长因子受体(EGFR)基因表达对食管鳞癌Eca109细胞放射敏感性的影响。方法 化学合成3种序列EGFR siRNA (EGFR siRNA1、EGFR siRNA2、EGFR siRNA3),设随机序列阳性siRNA组、随机序列阴性siRNA组、空白对照组,通过脂质体转染法转入Eca109细胞。以CCK8法检测细胞增殖,采用RT-PCR和Western blot检测干扰前后的Eca109细胞EGFR mRNA和蛋白表达。用细胞克隆形成实验分析EGFR siRNA联合X射线照射对Eca109细胞的放射敏感性影响。结果 EGFR siRNA1、EGFR siRNA2、EGFR siRNA3转染Eca109细胞后的EGFR mRNA的阳性表达率为26.74%、9.52%、4.61%,较空白对照组的42.44%明显下降(F=112.11,P<0.01),EGFR siRNA1、EGFR siRNA2、EGFR siRNA3对Eca109细胞EGFR mRNA表达抑制效率可高达72.84%、53.01%和56.21%;CCK8实验显示siRNA3干扰EGFR后Eca109细胞生长受到抑制,增殖抑制率为28.2%;成克隆分析的EGFR siRNA3 转染Eca109细胞加照射组D0、Dq、SF2值均低于单纯照射组,放射增敏比(SER)为1.50。结论 EGFR siRNA转染Eca109细胞后能有效抑制EGFR基因表达,提高其放射敏感性。 |
| 英文摘要: |
| Objective To explore the influence of EGFR gene interfering on the radiosensitivity of esophageal squamous cells.Methods Three kinds of siRNAs including three random sequences of positive EGFR siRNA (EGFR siRNA1、EGFR siRNA2、EGFR siRNA3), random sequence of negative EGFR siRNA, and blank control were transfected into Eca109 cells by lipofectamine. Cell proliferation was detected by Cell Counting Kit-8 assay. Protein and mRNA expressions of EGFR were detected by Western blot and RT-PCR, respectively. The ability of cell clone formation was used to evaluate the combination effect of X-rays and EGFR siRNA on the radiosensitivity.Results The positive expression rate of the EGFR mRNA in the Eca109 cells transfected with EGFR siRNA1, EGFR siRNA2, EGFR siRNA3 was 26.74%, 9.52%, 4.61%, respectively, which was significantly lower than 42.44% in the control cells transfected with blank siRNA(F=112.11,P<0.01). Meanwhile, the EGFR protein expression was reduced by 72.84%, 53.01% and 56.21% after interfering of siRNA1, siRNA2, and siRNA3, respectively. CCK8 assay showed that the proliferation of Eca109 cells was decreased by 28.2% since the siRNA interference. Moreover, the D0, Dq and SF2 of the combined treatment group were lower than those of irradiation alone group and the sensitization enhancement ratio was 1.50.Conclusions EGFR siRNA can effectively inhibit EGFR gene expression and enhance the radiosensitivity of Eca109 cells. |
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