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. 1993 Feb;67(2):279–283. doi: 10.1038/bjc.1993.53

Inhibition of human pancreatic cancer cell (MIA PaCa-2) growth by cholera toxin and 8-chloro-cAMP in vitro.

E Ohmura 1, K Wakai 1, O Isozaki 1, H Murakami 1, N Onoda 1, N Emoto 1, K Shizume 1, T Tsushima 1, H Demura 1, R K Robins 1
PMCID: PMC1968158  PMID: 8381655

Abstract

The effects of cholera toxin (CT) and 8-chloro-cAMP (8-Cl-cAMP) on cell growth were investigated using two human pancreatic carcinoma cell lines (MIA PaCa-2, Panc-1). CT, which catalyses the ADP ribosylation of Gs, suppresses the proliferation of MIA PaCa-2(PC) cells. CT at the low dose of 0.1 pg ml-1 was inhibitory of PC cell growth, and the maximum suppression (70%) was achieved at a CT concentration of 100 pg ml-1. This phenomenon was reversible. The production of cAMP by CT (100 pg ml-1) in PC cells was enhanced 320-fold compared with the control. In addition, cAMP analogues (8-Cl-cAMP, 8-Br-cAMP) and forskolin decreased the growth rate of PC cells in a dose-dependent manner. These results support the view that CT suppresses PC cell growth by stimulating cAMP production. Conversely, Panc-1 cells were far less sensitive to CT in cell growth and cAMP production. 8-Cl-cAMP was also less effective on Panc-1 cell growth. The binding of an insulin-like growth factor (IGF)-I and transforming growth factor (TGF)-alpha, which has been shown to stimulate PC cell growth in an autocrine manner, to PC cells was not modified in cells treated with CT or 8-Cl-cAMP. The results suggest that the inhibitory actions of these substances do not occur at the level of the receptor for IGF-I or EGF/TGF-alpha. We have previously shown that phorbol esters, which decrease the binding of TGF-alpha to PC cells, has an anti-proliferative activity on these tumour cells. Inhibited cell growth by maximum suppressive dose of CT or 8-Cl-cAMP was further inhibited by TPA. In addition, an oncogene product of K-ras which is commonly activated in pancreatic cancer, was increased by CT and 8-Cl-cAMP. It is concluded that CT and 8-Cl-cAMP inhibit PC cell growth, presumably in a similar manner, and their mechanism(s) of action may be different from that of TPA. The anti-proliferative effect of CT or 8-Cl-cAMP was enhanced by TPA, implying that the combination of these substances results in increased inhibition of the PC cell growth.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ally S., Clair T., Katsaros D., Tortora G., Yokozaki H., Finch R. A., Avery T. L., Cho-Chung Y. S. Inhibition of growth and modulation of gene expression in human lung carcinoma in athymic mice by site-selective 8-Cl-cyclic adenosine monophosphate. Cancer Res. 1989 Oct 15;49(20):5650–5655. [PubMed] [Google Scholar]
  2. Ally S., Tortora G., Clair T., Grieco D., Merlo G., Katsaros D., Ogreid D., Døskeland S. O., Jahnsen T., Cho-Chung Y. S. Selective modulation of protein kinase isozymes by the site-selective analog 8-chloroadenosine 3',5'-cyclic monophosphate provides a biological means for control of human colon cancer cell growth. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6319–6322. doi: 10.1073/pnas.85.17.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  4. Barbacid M. ras oncogenes: their role in neoplasia. Eur J Clin Invest. 1990 Jun;20(3):225–235. doi: 10.1111/j.1365-2362.1990.tb01848.x. [DOI] [PubMed] [Google Scholar]
  5. Chesa P. G., Rettig W. J., Melamed M. R., Old L. J., Niman H. L. Expression of p21ras in normal and malignant human tissues: lack of association with proliferation and malignancy. Proc Natl Acad Sci U S A. 1987 May;84(10):3234–3238. doi: 10.1073/pnas.84.10.3234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cho-Chung Y. S., Clair T., Shepheard C., Berghoffer B. Arrest of hormone-dependent mammary cancer growth in vivo and in vitro by cholera toxin. Cancer Res. 1983 Apr;43(4):1473–1476. [PubMed] [Google Scholar]
  7. Cho-Chung Y. S. Role of cyclic AMP receptor proteins in growth, differentiation, and suppression of malignancy: new approaches to therapy. Cancer Res. 1990 Nov 15;50(22):7093–7100. [PubMed] [Google Scholar]
  8. Cochet C., Gill G. N., Meisenhelder J., Cooper J. A., Hunter T. C-kinase phosphorylates the epidermal growth factor receptor and reduces its epidermal growth factor-stimulated tyrosine protein kinase activity. J Biol Chem. 1984 Feb 25;259(4):2553–2558. [PubMed] [Google Scholar]
  9. Feramisco J. R., Clark R., Wong G., Arnheim N., Milley R., McCormick F. Transient reversion of ras oncogene-induced cell transformation by antibodies specific for amino acid 12 of ras protein. Nature. 1985 Apr 18;314(6012):639–642. doi: 10.1038/314639a0. [DOI] [PubMed] [Google Scholar]
  10. Holmgren J. Actions of cholera toxin and the prevention and treatment of cholera. Nature. 1981 Jul 30;292(5822):413–417. doi: 10.1038/292413a0. [DOI] [PubMed] [Google Scholar]
  11. Howe P. H., Bascom C. C., Cunningham M. R., Leof E. B. Regulation of transforming growth factor beta 1 action by multiple transducing pathways: evidence for both G protein-dependent and -independent signaling. Cancer Res. 1989 Nov 1;49(21):6024–6031. [PubMed] [Google Scholar]
  12. Iwashita S., Fox C. F. Epidermal growth factor and potent phorbol tumor promoters induce epidermal growth factor receptor phosphorylation in a similar but distinctively different manner in human epidermoid carcinoma A431 cells. J Biol Chem. 1984 Feb 25;259(4):2559–2567. [PubMed] [Google Scholar]
  13. Lechner J. F., Haugen A., Autrup H., McClendon I. A., Trump B. F., Harris C. C. Clonal growth of epithelial cells from normal adult human bronchus. Cancer Res. 1981 Jun;41(6):2294–2304. [PubMed] [Google Scholar]
  14. Lee L. S., Weinstein I. B. Tumor-promoting phorbol esters inhibit binding of epidermal growth factor to cellular receptors. Science. 1978 Oct 20;202(4365):313–315. doi: 10.1126/science.308698. [DOI] [PubMed] [Google Scholar]
  15. Mukhopadhyay T., Tainsky M., Cavender A. C., Roth J. A. Specific inhibition of K-ras expression and tumorigenicity of lung cancer cells by antisense RNA. Cancer Res. 1991 Mar 15;51(6):1744–1748. [PubMed] [Google Scholar]
  16. Ohmura E., Okada M., Onoda N., Kamiya Y., Murakami H., Tsushima T., Shizume K. Insulin-like growth factor I and transforming growth factor alpha as autocrine growth factors in human pancreatic cancer cell growth. Cancer Res. 1990 Jan 1;50(1):103–107. [PubMed] [Google Scholar]
  17. Rozengurt E., Legg A., Strang G., Courtenay-Luck N. Cyclic AMP: a mitogenic signal for Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4392–4396. doi: 10.1073/pnas.78.7.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shoyab M., De Larco J. E., Todaro G. J. Biologically active phorbol esters specifically alter affinity of epidermal growth factor membrane receptors. Nature. 1979 May 31;279(5712):387–391. doi: 10.1038/279387a0. [DOI] [PubMed] [Google Scholar]
  19. Tagliaferri P., Katsaros D., Clair T., Ally S., Tortora G., Neckers L., Rubalcava B., Parandoosh Z., Chang Y. A., Revankar G. R. Synergistic inhibition of growth of breast and colon human cancer cell lines by site-selective cyclic AMP analogues. Cancer Res. 1988 Mar 15;48(6):1642–1650. [PubMed] [Google Scholar]
  20. Takanashi A., Yasui W., Yoshida K., Yokozaki H., Saito D., Abe K., Urakami K., Miki K., Tahara E. Inhibitory effect of 8-chloro-cyclic adenosine 3',5'-monophosphate on cell growth of gastric carcinoma cell lines. Jpn J Cancer Res. 1991 Mar;82(3):325–331. doi: 10.1111/j.1349-7006.1991.tb01849.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Taylor-Papadimitriou J., Purkis P., Fentiman I. S. Cholera toxin and analogues of cyclic AMP stimulate the growth of cultured human mammary epithelial cells. J Cell Physiol. 1980 Mar;102(3):317–321. doi: 10.1002/jcp.1041020306. [DOI] [PubMed] [Google Scholar]
  22. Tortora G., Ciardiello F., Ally S., Clair T., Salomon D. S., Cho-Chung Y. S. Site-selective 8-chloroadenosine 3',5'-cyclic monophosphate inhibits transformation and transforming growth factor alpha production in Ki-ras-transformed rat fibroblasts. FEBS Lett. 1989 Jan 2;242(2):363–367. doi: 10.1016/0014-5793(89)80502-2. [DOI] [PubMed] [Google Scholar]
  23. Tortora G., Tagliaferri P., Clair T., Colamonici O., Neckers L. M., Robins R. K., Cho-Chung Y. S. Site-selective cAMP analogs at micromolar concentrations induce growth arrest and differentiation of acute promyelocytic, chronic myelocytic, and acute lymphocytic human leukemia cell lines. Blood. 1988 Jan;71(1):230–233. [PubMed] [Google Scholar]
  24. Viallet J., Sharoni Y., Frucht H., Jensen R. T., Minna J. D., Sausville E. A. Cholera toxin inhibits signal transduction by several mitogens and the in vitro growth of human small-cell lung cancer. J Clin Invest. 1990 Dec;86(6):1904–1912. doi: 10.1172/JCI114923. [DOI] [PMC free article] [PubMed] [Google Scholar]

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