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Construction of eukaryotic expression plasmid expressing siRNA targeting ATM gene and its inhibitory effect on ATM expression in transfected human cervical carcinoma cells
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 5
- DOI:
- 10.3321/j.issn:1000-2790.2005.05.003
- Key Word:
- ATM基因;RNA干扰;宫颈癌;转染
Abstract： 目的:研究特异性siRNA对宫颈癌ATM基因的阻抑效果以及用RNAi效应对宫颈癌做基因治疗的可行性. 方法:构建可表达人ATM基因siRNA的重组真核表达质粒pSuppressorNeo-ATM,电穿孔法转染Siha宫颈癌细胞株. 应用RT-PCR,Western blot,流式细胞术、免疫荧光法等方法检测转染的宫颈癌细胞中ATM基因的表达水平. 结果:RT-PCR, Western-blot均表明瞬时转染pSuppressorNeo-ATM的宫颈癌细胞中ATM 基因的表达受到明显抑制,流式细胞术、免疫荧光法检测表明ATM蛋白含量明显下降. 结论:pSuppressorNeo-ATM质粒构建成功,瞬时转染宫颈癌细胞后可以明显抑制ATM基因的表达.
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