Johnson M D, Davis B W, Wetherall N T
Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN 37232.
J Environ Pathol Toxicol Oncol. 1989 Mar-Apr;9(2):171-90.
There is increasing interest in cellular localization of protooncogene, growth factor, and growth factor receptor gene expression in developing or neoplastic human tissue. Establishment of nucleic acid in situ hybridization (ISH) and immunohistochemical (IH) analyses of these genes, however, is compromised by the paucity of available control tissues which have been shown to express these cancer associated genes. The present study evaluated whether proto-oncogene or growth factor gene expressing human control tissues suitable for ISH and IH studies could be produced from malignant cell lines implanted into irradiated nude mice. Tumors rapidly fixed in parafomaldehyde 2 to 4 weeks after xenograft were found optimal for in situ hybridization and immunohistochemistry. Northern blots of frozen tumors confirmed the expression of specific proto-oncogenes in the tissue and the specificity of cDNA and cRNA probes for those transcripts. Production of control tissue from xenografted cell lines should facilitate establishment and long term quality control of in situ hybridization and immunohistochemical analysis of proto-oncogene or growth factor gene expression in laboratories with limited experience in these techniques.
人们对原癌基因、生长因子和生长因子受体基因在发育中的或肿瘤性人类组织中的细胞定位的兴趣与日俱增。然而，由于缺乏已被证明表达这些癌症相关基因的可用对照组织，这些基因的核酸原位杂交（ISH）和免疫组织化学（IH）分析的建立受到了影响。本研究评估了植入经辐射的裸鼠体内的恶性细胞系能否产生适合ISH和IH研究的表达原癌基因或生长因子基因的人类对照组织。异种移植后2至4周迅速固定在多聚甲醛中的肿瘤被发现最适合进行原位杂交和免疫组织化学检测。冷冻肿瘤的Northern印迹证实了组织中特定原癌基因的表达以及cDNA和cRNA探针针对这些转录本的特异性。从异种移植细胞系产生对照组织应有助于在这些技术方面经验有限的实验室建立原位杂交和免疫组织化学分析，并进行长期质量控制，以检测原癌基因或生长因子基因的表达。
