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肿瘤细胞由多聚甲醛直接固定在硝酸纤维素膜上,经蛋白酶消化后与地高辛配基(di-goxigenin)标记的癌基因探针进行DNA-RNA杂交,洗膜后用酶联免疫法显色,可以半定量的确定癌基因在肿瘤细胞中的表达状况。该方法不需提取RNA,因此操作简便易行。由于利用地高辛配基标记探针,克服了同位素放射性和蜕变带来的诸多不便。实验将此法用于肺癌细胞经诱导分化后癌基因表达改变以及白血病细胞中癌基因表达情况研究,5×10~3个细胞即可获得阳性杂交信号。
Tumor cells were fixed directly on nitrocellulose membrane by paraformaldehyde, then digested with di-goxigenin, digested with protease and subjected to DNA-RNA hybridization. Color, semi-quantitative determination of cancer gene expression in tumor cells. This method does not need to extract RNA, so the operation is simple and easy. Due to the use of digoxigenin-labeled probes, overcoming the inconveniences caused by radioactive isotopes and metamorphosis. This method was used to study the change of oncogene expression in lung cancer cells induced by differentiation and the expression of oncogene in leukemia cells. The positive hybridization signal was obtained in 5 × 10 ~ 3 cells.