摘要
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 method of non-isotope double in situ hybridization in order to detect the expression of two kinds of oncogenes at a single cell level simultaneously,and confirm the hypothesis of 'model of stepwise carcinogenesis'.Methods:The method of non-isotope doulbe in situ hybridization was established with the digoxigenin(Dig) and biotint(Bio) labeled probes.The expression of two members of oncogenes of the myc and/or ras gene families(myc and N-ras,myc and K-ras,myc and H-ras ,N-ras and K-ras, N-ras and H-ras,K-ras and H-ras) was further studied with the method in 10 cases of Chinese Hepatocellular Carcinomas(HCC).Results:Co-expression of two kinds of oncogenes was detected only in a few cases,reflected by co-showing two different hybridization signals,i. e.,co-showing of Dig-myc and Bio-H-ras,Dig-myc and Eio-N-ras , Dig-myc and Bio-K-ras was observed in 2 cases , 2 cases and one case ,respectively. The common characteristics were that positive cells of myc distributed diffusely, while positive cells of ras distributed sporadically or locally among the positive cells of myc,and only a few cells exibited co-showing of two oncogenes at a single cell level. There were only 2 cases representing co-showing of two oncogenes in ras gene family (Dig-N-ras and Bio-H-ras , Dig-N-ras and Bio-Kras,respectively).The two kinds of positive cells of different ras gene represented mixed local andsporadical distribution.and co-showing of two signals was found in a few cells at a single cell level. Conclusion: There are multiple oncogenes involving in tumorigenesis by their ordered activation. The activation of the ras gene family plays a role in promotion,while the activation of myc is an important event in late stage of tumorigenesis.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
1996年第1期6-9,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金
