小鼠编码锌指蛋白基因zfp637的初步研究

Primary Study of Mouse Zinc Finger Gene zfp637

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144d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，143d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。

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 144. 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 peripheral blood leukocyte. The zfp637-specific siRNAs substantially reduced the proliferation of EMT6 cells on the 4th day. Conclusion zfp637 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 its expression in EMT6 cancer cells by RNAi led to a remarkable reduction of proliferation.