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Detection of human probombesin mrna in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with crna probe
Author(s) -
Hamid Qutayba A.,
Bishop Anne E.,
Springall David R.,
Adams Caroline,
Giaid Adel,
Denny Paul,
Ghatei Mohammed,
Legon Stephen,
Cuttitta Frank,
Rode Jurgen,
Spindel Eliot,
Bloom Stephen R.,
Polak Julia M.
Publication year - 1989
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/1097-0142(19890115)63:2<266::aid-cncr2820630211>3.0.co;2-d
Subject(s) - bombesin , in situ hybridization , microbiology and biotechnology , biology , small cell carcinoma , gene expression , complementary dna , carcinoma , immunostaining , messenger rna , gene , immunohistochemistry , neuropeptide , immunology , receptor , biochemistry , genetics
The production of human bombesin (gastrin‐releasing peptide), a peptide with mitogenic action, is a recognized feature of neuroendocrine (small cell) carcinoma of the lung. However, immunostaining of bombesin is not always possible in these tumors, probably because of poor storage mechanisms or rapid release of hormone. Molecular biological analysis of the gene encoding human bombesin has revealed the DNA sequence of human pro‐bombesin. We have used in situ hybridization to study the expression of the human bombesin gene at the cellular level in small cell carcinoma of the lung. Probombesin cDNA was subcloned in pSP64 vector, linearized with Bam HI and transcribed in the presence of phosphorus 32 ( 32 P)‐cytosine triphosphate (CTP) and SP6 polymerase. The cRNA probe was applied to tissue sections (from six cases of small cell carcinoma of the lung, freshly fixed in 4% paraformaldehyde), cell culture preparations (two different cell lines of small cell carcinoma), and cytologic specimens (smears of cells from three different cases of small cell carcinoma). Hybridization of probombesin mRNA was detected in tumor cells in all samples. Specificity of the signal was determined by control experiments, including the use of a probe which has a sequence identical to probombesin mRNA. Our results provide evidence for the expression of the bombesin gene in small cell carcinoma of the lung at a cellular level and show that probombesin mRNA is highly expressed in these tumors.