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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。
Objective To observe the effect of zfp637 gene on tumor cell growth and proliferation. Methods Semi-quantitative RT-PCR was used to detect the expression of zfp637 gene in mouse normal tissues and tumor cell lines. Four pairs of small double-stranded RNAs were designed and synthesized, and the best interference effect sites were screened by semi-quantitative RT-PCR. The siRNA was transfected into EMT6 cells and cell proliferation experiments were performed 1 ~ 4 days after transfection. Results zfp637 showed low to moderate expression in most normal tissues and no expression of peripheral blood mononuclear cells. High expression was found in mouse liver cancer H22, mouse hepatoma Hepal-6, Lewis lung cancer LL / 2, melanoma B16, lymphoma Yac-1 and breast cancer EMT6. After screening, it showed that siRNA-881 was the best interference effect site. Cell proliferation assay (MTT) showed that the proliferation of EMT6 cells in the experimental group was significantly lower than that in the negative control group on the 4th day after transfection, and the difference between the 1 and 3d experimental groups was not obvious. Conclusion zfp637 gene expression in different normal tissues and tumor cell lines in different levels. zfp637 is likely to be a positive regulator of cell proliferation. Down-regulation of the gene expression can inhibit the proliferation of EMT6 tumor cells.