目的：建立非同位素双重原位杂交技术，以便在单细胞水平同时显示两种不同的癌基因杂交信号，验证＂癌变多步骤＂学说。方法：采用地高辛（Ｄｉｇ）和生物素（Ｂｉｏ）标记探针，成功地建立了非同位素双重原位杂交技术，对１０例人肝细胞性肝癌（ＨＣＣ）及癌周组织细胞内ｍｙｃ、ｒａｓ癌基因家族及ｒａｓ家族内部基因两－两之间（ｍｙｃ与Ｎ－ｒａｓ，ｍｙｃ与Ｋ－ｒａｓ，ｍｙｃ与Ｈ－ｒａｓ，Ｎ－ｒａｓ与Ｋ－ｒａｓ，Ｎ－ｒａｓ与Ｈ－ｒａｓ，Ｋ－ｒａｓ与Ｈ－ｒａｓ）的表达情况进行了检测。结果：仅在个别肿瘤标本中少量肿瘤细胞内发现有两种癌基因杂交信号的共同显示。Ｄｉｇ－ｍｙｃ与Ｂｉｏ－Ｈ－ｒａｓ共同显示２例，Ｄｉｇ－ｍｙｃ与Ｂｉｏ－Ｎ－ｒａｓ共同显示２例，Ｄｉｇ－ｍｙｃ与Ｂｉｏ－Ｋ－ｒａｓ共同显示１例，其分布共同特点是ｍｙｃ阳性细胞呈弥漫性分布，ｒａｓ阳性细胞呈散在或片状分布于ｍｙｃ阳性细胞之间，偶见少量细胞内有两种核酸杂交信号的共同显示；ｒａｓ家族中Ｄｉｇ－Ｎ－ｒａｓ与Ｂｉｏ－Ｈ－ｒａｓ、Ｄｉｇ－Ｎ－ｒａｓ与Ｂｉｏ－Ｋ－ｒａｓ仅各１例共同显示。两种阳性细胞各呈片状、散在性分布，有部分混杂，其中少量细胞内有两种核酸杂交信号的共同显示。结论：? OBJECTIVE: To establish a non-isotopic dual in situ hybridization technique to simultaneously display two different oncogene hybridization signals at the single cell level and to verify the “multistep cancer” doctrine. METHODS: Digoxin (Dig) and biotin (Bio) labeled probes were used to successfully establish a non-isotopic dual in situ hybridization technique for intracellular myc in 10 cases of human hepatocellular carcinoma (HCC) and pericancerous tissue. The ras oncogene family and the internal genes of the ras family between two and two (myc and N-ras, myc and K-ras, myc and H-ras, N-ras and K-ras, N-ras and H-ras, K The expression of -ras and H-ras) was examined. RESULTS: A common display of two oncogene hybridization signals was found only in a small number of tumor cells in individual tumor specimens. Two cases of Dig-myc and Bio-H-ras were shown together. Two cases of Dig-myc and Bio-N-ras were shown together. One case of Dig-myc and Bio-K-ras was shown together. Their distribution was characterized by myc-positive cells. Diffusely distributed, ras positive cells are scattered or flaky distributed between myc positive cells. Occasionally, a few cells show the hybridization signals of two kinds of nucleic acids; Digs-N-ras and Bio-H-ras in ras family. Only one case of Dig-N-ras and Bio-K-ras was displayed. The two positive cells showed a patchy and scattered distribution, some of which were mixed, and a small number of cells showed the hybridization signals of the two nucleic acids. in conclusion:?