Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with cRNA probe

Q. A. Hamid, A. E. Bishop, D. R. Springall, C. Adams, A. Giaid, P. Denny, M. Ghatei, S. Legon, F. Cuttitta, J. Rode, Eliot Spindel, S. R. Bloom, J. M. Polak

Research output: Contribution to journalArticle

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Abstract

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 (32P)-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.

Original languageEnglish (US)
Pages (from-to)266-271
Number of pages6
JournalCancer
Volume63
Issue number2
StatePublished - 1989
Externally publishedYes

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Bombesin
Neuroendocrine Cells
Complementary RNA
Small Cell Carcinoma
In Situ Hybridization
Lung
Messenger RNA
Small Cell Lung Carcinoma
Gastrin-Releasing Peptide
Neoplasms
Cytosine
Phosphorus
Genes
Complementary DNA
Cell Culture Techniques
Hormones
Gene Expression
Cell Line
Peptides

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Hamid, Q. A., Bishop, A. E., Springall, D. R., Adams, C., Giaid, A., Denny, P., ... Polak, J. M. (1989). Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with cRNA probe. Cancer, 63(2), 266-271.

Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with cRNA probe. / Hamid, Q. A.; Bishop, A. E.; Springall, D. R.; Adams, C.; Giaid, A.; Denny, P.; Ghatei, M.; Legon, S.; Cuttitta, F.; Rode, J.; Spindel, Eliot; Bloom, S. R.; Polak, J. M.

In: Cancer, Vol. 63, No. 2, 1989, p. 266-271.

Research output: Contribution to journalArticle

Hamid, QA, Bishop, AE, Springall, DR, Adams, C, Giaid, A, Denny, P, Ghatei, M, Legon, S, Cuttitta, F, Rode, J, Spindel, E, Bloom, SR & Polak, JM 1989, 'Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with cRNA probe', Cancer, vol. 63, no. 2, pp. 266-271.
Hamid QA, Bishop AE, Springall DR, Adams C, Giaid A, Denny P et al. Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with cRNA probe. Cancer. 1989;63(2):266-271.
Hamid, Q. A. ; Bishop, A. E. ; Springall, D. R. ; Adams, C. ; Giaid, A. ; Denny, P. ; Ghatei, M. ; Legon, S. ; Cuttitta, F. ; Rode, J. ; Spindel, Eliot ; Bloom, S. R. ; Polak, J. M. / Detection of human probombesin mRNA in neuroendocrine (small cell) carcinoma of the lung. In situ hybridization with cRNA probe. In: Cancer. 1989 ; Vol. 63, No. 2. pp. 266-271.
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abstract = "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 (32P)-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.",
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AU - Hamid, Q. A.

AU - Bishop, A. E.

AU - Springall, D. R.

AU - Adams, C.

AU - Giaid, A.

AU - Denny, P.

AU - Ghatei, M.

AU - Legon, S.

AU - Cuttitta, F.

AU - Rode, J.

AU - Spindel, Eliot

AU - Bloom, S. R.

AU - Polak, J. M.

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