小鼠编码锌指蛋白基因zfp637的初步研究
Primary Study of Mouse Zinc Finger Gene zfp637
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摘要: 目的观察zfp637基因对肿瘤细胞生长增殖的影响。方法半定量RT-PCR检测zfp637基因在小鼠正常组织与肿瘤细胞株中的表达。设计并合成4对小分子双链RNA,半定量RT-PCR筛选最佳干扰效应位点。将siRNA转染EMT6细胞,转染后1~4d进行细胞增殖实验。结果zfp637在多数正常组织呈低到中度表达,外周血单个核细胞未见表达。在小鼠肝癌H22,小鼠肝癌细胞Hepal-6,Lewis肺癌LL/2,黑色素瘤细胞B16,淋巴瘤细胞Yac-1和乳腺癌细胞EMT6中均呈现高表达。筛选后表明siRNA-881为最佳干扰效应位点。细胞增殖实验(MTT)显示:转染siRNA后第4d,实验组EMT6细胞增殖明显低于阴性对照组,1~3d实验组细胞增殖与阴性组相差不明显。结论zfp637基因在不同正常组织和肿瘤细胞株中表达水平不同。zfp637很可能是促进细胞增殖的正调控因素。下调该基因表达能抑制EMT6肿瘤细胞增殖。Abstract: Objective To investigate the impact of zfp637 on proliferation of cancer cells. Methods The expression of zfp637 in normal tissues and cancer cell lines was examined by semi-quantity RT-PCR analysis.Small double strands interfering RNA(siRNA) was synthesized,and then screen the best interfering site.After that,proliferation of RNAi treated EMT6 cells was analyzed via 96-well plate approach.The assay was performed on day 1~4. Results zfp637 was expressed moderately in most normal tissues,highly expressed in Yac-1, EMT6, B16, H22, LL/ 2 and Hepal-6, but not expressed in pe-ripheral blood leukocyte. The z f p637-specific siRNAs substantially reduced the proliferation of EMT6 cells on the 4th day. Conclusion z f p637 was expressed at different levels in multiple tissues and various cancer cell lines. It probably functions as a positive regulator of cellular proliferation. Down regulation of it s expression in EMT6 cancer cells by RNAi led to a remarkable reduction of proliferation.
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