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. 1999 Aug;80(12):1935–1944. doi: 10.1038/sj.bjc.6690623

Tumour-specific arginine vasopressin promoter activation in small-cell lung cancer

J M Coulson 1, J Stanley 1, P J Woll 1
PMCID: PMC2374275  PMID: 10471042

Abstract

Small-cell lung cancer (SCLC) can produce numerous mitogenic neuropeptides, which are not found in normal respiratory epithelium. Arginine vasopressin is detected in up to two-thirds of SCLC tumours whereas normal physiological expression is essentially restricted to the hypothalamus. This presents the opportunity to identify elements of the gene promoter which could be exploited for SCLC-specific targeting. A series of human vasopressin 5′ promoter fragments (1048 bp, 468 bp and 199 bp) were isolated and cloned upstream of a reporter gene. These were transfected into a panel of ten cell lines, including SCLC with high or low endogenous vasopressin transcription, non-SCLC and bronchial epithelium. All these fragments directed reporter gene expression in the five SCLC cell lines, but had negligible activity in the control lines. The level of reporter gene expression reflected the level of endogenous vasopressin production, with up to 4.9-fold (s.d. 0.34) higher activity than an SV40 promoter. The elements required for this strong, restricted, SCLC-specific promoter activity are contained within the 199-bp fragment. Further analysis of this region indicated involvement of E-box transcription factor binding sites, although tumour-specificity was retained by a 65-bp minimal promoter fragment. These data show that a short region of the vasopressin promoter will drive strong expression in SCLC in vitro and raise the possibility of targeting gene therapy to these tumours. © 1999 Cancer Research Campaign

Keywords: vasopressin, promoter, SCLC, transcription, tumour-specific, enhancer

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Selected References

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  1. Ang H. L., Carter D. A., Murphy D. Neuron-specific expression and physiological regulation of bovine vasopressin transgenes in mice. EMBO J. 1993 Jun;12(6):2397–2409. doi: 10.1002/j.1460-2075.1993.tb05894.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bahnsen U., Oosting P., Swaab D. F., Nahke P., Richter D., Schmale H. A missense mutation in the vasopressin-neurophysin precursor gene cosegregates with human autosomal dominant neurohypophyseal diabetes insipidus. EMBO J. 1992 Jan;11(1):19–23. doi: 10.1002/j.1460-2075.1992.tb05022.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ball D. W., Azzoli C. G., Baylin S. B., Chi D., Dou S., Donis-Keller H., Cumaraswamy A., Borges M., Nelkin B. D. Identification of a human achaete-scute homolog highly expressed in neuroendocrine tumors. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5648–5652. doi: 10.1073/pnas.90.12.5648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baum C., Forster P., Hegewisch-Becker S., Harbers K. An optimized electroporation protocol applicable to a wide range of cell lines. Biotechniques. 1994 Dec;17(6):1058–1062. [PubMed] [Google Scholar]
  5. Burke Z. D., Ho M. Y., Morgan H., Smith M., Murphy D., Carter D. Repression of vasopressin gene expression by glucocorticoids in transgenic mice: evidence of a direct mechanism mediated by proximal 5' flanking sequence. Neuroscience. 1997 Jun;78(4):1177–1185. doi: 10.1016/s0306-4522(96)00603-3. [DOI] [PubMed] [Google Scholar]
  6. Fay M. J., Friedmann A. S., Yu X. M., North W. G. Vasopressin and vasopressin-receptor immunoreactivity in small-cell lung carcinoma (SCCL) cell lines: disruption in the activation cascade of V1a-receptors in variant SCCL. Cancer Lett. 1994 Jul 29;82(2):167–174. doi: 10.1016/0304-3835(94)90007-8. [DOI] [PubMed] [Google Scholar]
  7. Fiering S. N., Roederer M., Nolan G. P., Micklem D. R., Parks D. R., Herzenberg L. A. Improved FACS-Gal: flow cytometric analysis and sorting of viable eukaryotic cells expressing reporter gene constructs. Cytometry. 1991;12(4):291–301. doi: 10.1002/cyto.990120402. [DOI] [PubMed] [Google Scholar]
  8. Friedmann A. S., Malott K. A., Memoli V. A., Pai S. I., Yu X. M., North W. G. Products of vasopressin gene expression in small-cell carcinoma of the lung. Br J Cancer. 1994 Feb;69(2):260–263. doi: 10.1038/bjc.1994.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedmann A. S., Memoli V. A., North W. G. Vasopressin and oxytocin production by non-neuroendocrine lung carcinomas: an apparent low incidence of gene expression. Cancer Lett. 1993 Dec 10;75(2):79–85. doi: 10.1016/0304-3835(93)90191-b. [DOI] [PubMed] [Google Scholar]
  10. Grace C. O., Fink G., Quinn J. P. Characterization of potential regulatory elements within the rat arginine vasopressin proximal promoter. Neuropeptides. 1999 Feb;33(1):81–90. doi: 10.1054/npep.1999.0018. [DOI] [PubMed] [Google Scholar]
  11. Gross A. J., Steinberg S. M., Reilly J. G., Bliss D. P., Jr, Brennan J., Le P. T., Simmons A., Phelps R., Mulshine J. L., Ihde D. C. Atrial natriuretic factor and arginine vasopressin production in tumor cell lines from patients with lung cancer and their relationship to serum sodium. Cancer Res. 1993 Jan 1;53(1):67–74. [PubMed] [Google Scholar]
  12. Hara Y., Battey J., Gainer H. Structure of mouse vasopressin and oxytocin genes. Brain Res Mol Brain Res. 1990 Oct;8(4):319–324. doi: 10.1016/0169-328x(90)90045-f. [DOI] [PubMed] [Google Scholar]
  13. Huber B. E., Richards C. A., Krenitsky T. A. Retroviral-mediated gene therapy for the treatment of hepatocellular carcinoma: an innovative approach for cancer therapy. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8039–8043. doi: 10.1073/pnas.88.18.8039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Iwasaki Y., Oiso Y., Saito H., Majzoub J. A. Positive and negative regulation of the rat vasopressin gene promoter. Endocrinology. 1997 Dec;138(12):5266–5274. doi: 10.1210/endo.138.12.5639. [DOI] [PubMed] [Google Scholar]
  15. Jin X., Shearman L. P., Weaver D. R., Zylka M. J., de Vries G. J., Reppert S. M. A molecular mechanism regulating rhythmic output from the suprachiasmatic circadian clock. Cell. 1999 Jan 8;96(1):57–68. doi: 10.1016/s0092-8674(00)80959-9. [DOI] [PubMed] [Google Scholar]
  16. Johnson B. E., Chute J. P., Rushin J., Williams J., Le P. T., Venzon D., Richardson G. E. A prospective study of patients with lung cancer and hyponatremia of malignancy. Am J Respir Crit Care Med. 1997 Nov;156(5):1669–1678. doi: 10.1164/ajrccm.156.5.96-10075. [DOI] [PubMed] [Google Scholar]
  17. Kim J. K., Summer S. N., Wood W. M., Schrier R. W. Osmotic and non-osmotic regulation of arginine vasopressin (AVP) release, mRNA, and promoter activity in small cell lung carcinoma (SCLC) cells. Mol Cell Endocrinol. 1996 Oct 30;123(2):179–186. doi: 10.1016/s0303-7207(96)03912-3. [DOI] [PubMed] [Google Scholar]
  18. Kumagai T., Tanio Y., Osaki T., Hosoe S., Tachibana I., Ueno K., Kijima T., Horai T., Kishimoto T. Eradication of Myc-overexpressing small cell lung cancer cells transfected with herpes simplex virus thymidine kinase gene containing Myc-Max response elements. Cancer Res. 1996 Jan 15;56(2):354–358. [PubMed] [Google Scholar]
  19. Maier J. A., Voulalas P., Roeder D., Maciag T. Extension of the life-span of human endothelial cells by an interleukin-1 alpha antisense oligomer. Science. 1990 Sep 28;249(4976):1570–1574. doi: 10.1126/science.2218499. [DOI] [PubMed] [Google Scholar]
  20. Maurer L. H., O'Donnell J. F., Kennedy S., Faulkner C. S., Rist K., North W. G. Human neurophysins in carcinoma of the lung: relation to histology, disease stage, response rate, survival, and syndrome of inappropriate antidiuretic hormone secretion. Cancer Treat Rep. 1983 Nov;67(11):971–976. [PubMed] [Google Scholar]
  21. Murphy D., Funkhouser J., Ang H. L., Foo N. C., Carter D. Extrahypothalamic expression of the vasopressin and oxytocin genes. Ann N Y Acad Sci. 1993 Jul 22;689:91–106. doi: 10.1111/j.1749-6632.1993.tb55540.x. [DOI] [PubMed] [Google Scholar]
  22. North W. G., Fay M. J., Longo K. A., Du J. Expression of all known vasopressin receptor subtypes by small cell tumors implies a multifaceted role for this neuropeptide. Cancer Res. 1998 May 1;58(9):1866–1871. [PubMed] [Google Scholar]
  23. Okazaki T., Ishikawa T., Nishimori S., Igarashi T., Hata K., Fujita T. Hyperosmolarity-induced gene stimulation is mediated by the negative calcium responsive element. J Biol Chem. 1997 Dec 19;272(51):32274–32279. doi: 10.1074/jbc.272.51.32274. [DOI] [PubMed] [Google Scholar]
  24. Pardy K., Adan R. A., Carter D. A., Seah V., Burbach J. P., Murphy D. The identification of a cis-acting element involved in cyclic 3',5'-adenosine monophosphate regulation of bovine vasopressin gene expression. J Biol Chem. 1992 Oct 25;267(30):21746–21752. [PubMed] [Google Scholar]
  25. Pass M. K., Quintini G., Zarn J. A., Zimmermann S. M., Sigrist J. A., Stahel R. A. The 5'-flanking region of human CD24 gene has cell-type-specific promoter activity in small-cell lung cancer. Int J Cancer. 1998 Nov 9;78(4):496–502. doi: 10.1002/(sici)1097-0215(19981109)78:4<496::aid-ijc17>3.0.co;2-4. [DOI] [PubMed] [Google Scholar]
  26. Prestridge D. S. Predicting Pol II promoter sequences using transcription factor binding sites. J Mol Biol. 1995 Jun 23;249(5):923–932. doi: 10.1006/jmbi.1995.0349. [DOI] [PubMed] [Google Scholar]
  27. Prestridge D. S. SIGNAL SCAN 4.0: additional databases and sequence formats. Comput Appl Biosci. 1996 Apr;12(2):157–160. doi: 10.1093/bioinformatics/12.2.157. [DOI] [PubMed] [Google Scholar]
  28. Quinn J. P. Neuronal-specific gene expression--the interaction of both positive and negative transcriptional regulators. Prog Neurobiol. 1996 Nov;50(4):363–379. doi: 10.1016/s0301-0082(96)00041-x. [DOI] [PubMed] [Google Scholar]
  29. Sausville E., Carney D., Battey J. The human vasopressin gene is linked to the oxytocin gene and is selectively expressed in a cultured lung cancer cell line. J Biol Chem. 1985 Aug 25;260(18):10236–10241. [PubMed] [Google Scholar]
  30. Sethi T., Rozengurt E. Multiple neuropeptides stimulate clonal growth of small cell lung cancer: effects of bradykinin, vasopressin, cholecystokinin, galanin, and neurotensin. Cancer Res. 1991 Jul 1;51(13):3621–3623. [PubMed] [Google Scholar]
  31. Terasaki T., Matsuno Y., Shimosato Y., Yamaguchi K., Ichinose H., Nagatsu T., Kato K. Establishment of a human small cell lung cancer cell line producing a large amount of anti-diuretic hormone. Jpn J Cancer Res. 1994 Jul;85(7):718–722. doi: 10.1111/j.1349-7006.1994.tb02420.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Verbeeck M. A., Elands J. P., de Leij L. F., Buys C. H., Carney D. N., Bepler G., Roebroeck A. J., Van de Ven W. J., Burbach J. P. Expression of the vasopressin and gastrin-releasing peptide genes in small cell lung carcinoma cell lines. Pathobiology. 1992;60(3):136–142. doi: 10.1159/000163712. [DOI] [PubMed] [Google Scholar]
  33. Verbeeck M. A., Sutanto W., Burbach J. P. Regulation of vasopressin messenger RNA levels in the small cell lung carcinoma cell line GLC-8: interactions between glucocorticoids and second messengers. Mol Endocrinol. 1991 Jun;5(6):795–801. doi: 10.1210/mend-5-6-795. [DOI] [PubMed] [Google Scholar]
  34. Vile R. G., Hart I. R. Use of tissue-specific expression of the herpes simplex virus thymidine kinase gene to inhibit growth of established murine melanomas following direct intratumoral injection of DNA. Cancer Res. 1993 Sep 1;53(17):3860–3864. [PubMed] [Google Scholar]
  35. Yoon S. O., Chikaraishi D. M. Tissue-specific transcription of the rat tyrosine hydroxylase gene requires synergy between an AP-1 motif and an overlapping E box-containing dyad. Neuron. 1992 Jul;9(1):55–67. doi: 10.1016/0896-6273(92)90220-8. [DOI] [PubMed] [Google Scholar]

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