地高辛标记DNA探针快速筛选肿瘤细胞株基因表达差异

Detection of gene expression difference in tumor cell lines rapidly with DNA probe-labeled by Digoxin

目的建立寡核苷酸探针正向斑点杂交技术,快速筛选不同细胞株同一基因HIF-1a表达差异,探讨斑点杂交筛选基因表达的可行性。方法将多种培养肿瘤细胞株提取总RNA后,点样于尼龙膜上,与地高辛标记的HIF-1a基因探针杂交,结果与半定量RT-PCR结果比较。结果斑点杂交实验结果与RT-PCR结果一致,无统计学差异(P>0.05)。结论用随机引物地高辛标记DNA探针可以快速筛查不同细胞株同一基因表达差异,可以作为研究肿瘤基因表达差异的工具。

Objective To estabilish dot blot hybridization technology in the detection of HIF-1a gene expression of various tumor cells and to explore the availability of dot blot in screening gene expression difference.Methods Nuclear acid were extracted from a variety of cultured tumor cell lines and spotted on the nylon membrane,dot blot analysis and semi-quantitative RT-PCR were carded out respectively to detect HIF-1a gene expression.Results The dot-blot hybridization results are consistent with the RT-PCR.it did not reach the conventional level of statistical significance（P〈0.05）.Conclusion Dot blot hybridization with DIG-labeled HIF-1a probe can be used to screen quickly gene expressing difference in tumor cells and serve as a research tool for tumor gene expression difference.