Production of proto-oncogene expressing control tissues for in situ hybridization and immunohistochemical studies.
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.
Johnson
M D
MD
Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN 37232.
Davis
B W
BW
Wetherall
N T
NT
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015342:DNA Probes; D066246:ErbB Receptors / Q000235:genetics; D005786:Gene Expression Regulation; D006133:Growth Substances / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000235:genetics; D009693:Nucleic Acid Hybridization; D000090063:Proto-Oncogene Mas; D011519:Proto-Oncogenes; D014407:Tumor Cells, Cultured / Q000378:metabolism
D015342:DNA Probes; D006133:Growth Substances; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D066246:ErbB Receptors