Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1999 Jul;80(9):1350–1358. doi: 10.1038/sj.bjc.6690528

Inhibition of proliferation and induction of apoptosis in soft tissue sarcoma cells by interferon-α and retinoids

T Brodowicz 1, C Wiltschke 1, D Kandioler-Eckersberger 3, T W Grunt 1, M Rudas 4, S M Schneider 1, M Hejna 1, A Budinsky 1, C C Zielinski 1,2,5
PMCID: PMC2363080  PMID: 10424735

Abstract

Uncontrolled proliferation and a defect of apoptosis constitute crucial elements in the development and progression of tumours. Among many other biological response modifiers known to influence these mechanisms, the efficacy of retinoids and interferons in the treatment of various malignant entities is currently matter of discussion. In the present study, we have investigated the effects of 9-cis-retinoic acid (9cRA), 13-cis-retinoic acid (13cRA), all-trans-retinoic acid (tRA) and interferon-α on proliferation and apoptosis of human soft tissue sarcoma (STS) cell lines HTB-82 (rhabdomyosarcoma), HTB-91 (fibrosarcoma), HTB-92 (liposarcoma), HTB-93 (synovial sarcoma) and HTB-94 (chondrosarcoma) in relation to p53 genotype as well as p53 expression. HTB-91, HTB-92 and HTB-94 STS cells exhibited mutant p53, whereas wild-type p53 was found in HTB-93 STS cells, and a normal p53 status in HTB-82 STS cells, carrying a silent point mutation only. Interferon-α, irrespective of p53 status, inhibited the proliferation of all five cell lines dose- and time-dependently. Similarly, 9cRA, 13cRA and tRA decreased the proliferation of HTB-82 and HTB-93 STS cells, whereas the proliferation of p53-mutated HTB-91, HTB-92 and HTB-94 STS cells remained unchanged. Furthermore, only 9cRA and tRA were capable of inducing apoptosis in HTB-82 and HTB-93 STS cells, whereas HTB-91, HTB-92 and HTB-94 STS cells did not undergo apoptosis under the influence of 9cRA or tRA. Retinoic acid receptor (RAR)-α and RAR-β mRNA were not detectable by Northern blot analysis in the five STS cell lines, whereas mRNA for the universal retinoic acid receptor, RAR-γ, was expressed in all STS cell lines indicating that retinoid resistance was not associated with a lack of RAR expression. Apoptosis was not induced by interferon-α or 13cRA in any of the five STS cell lines tested. Our results indicate that within the panel of tested STS cell lines, inhibition of proliferation and induction of apoptosis result from different mechanisms which differ in their dependence upon the presence of intact p53. © 1999 Cancer Research Campaign

Keywords: apoptosis, soft tissue sarcoma, interferon-α, retinoids, p53

Full Text

The Full Text of this article is available as a PDF (365.4 KB).

Selected References

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

  1. Aas T., Børresen A. L., Geisler S., Smith-Sørensen B., Johnsen H., Varhaug J. E., Akslen L. A., Lønning P. E. Specific P53 mutations are associated with de novo resistance to doxorubicin in breast cancer patients. Nat Med. 1996 Jul;2(7):811–814. doi: 10.1038/nm0796-811. [DOI] [PubMed] [Google Scholar]
  2. Agarwal M. L., Agarwal A., Taylor W. R., Stark G. R. p53 controls both the G2/M and the G1 cell cycle checkpoints and mediates reversible growth arrest in human fibroblasts. Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8493–8497. doi: 10.1073/pnas.92.18.8493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Antman K. H. Adjuvant therapy of sarcomas of soft tissue. Semin Oncol. 1997 Oct;24(5):556–560. [PubMed] [Google Scholar]
  4. Antman K., Crowley J., Balcerzak S. P., Rivkin S. E., Weiss G. R., Elias A., Natale R. B., Cooper R. M., Barlogie B., Trump D. L. An intergroup phase III randomized study of doxorubicin and dacarbazine with or without ifosfamide and mesna in advanced soft tissue and bone sarcomas. J Clin Oncol. 1993 Jul;11(7):1276–1285. doi: 10.1200/JCO.1993.11.7.1276. [DOI] [PubMed] [Google Scholar]
  5. Berggren Söderlund M., Johannesson G., Fex G. Expression of human all-trans-retinoic acid receptor beta and its ligand-binding domain in Escherichia coli. Biochem J. 1995 May 15;308(Pt 1):353–359. [PMC free article] [PubMed] [Google Scholar]
  6. Bergh J., Norberg T., Sjögren S., Lindgren A., Holmberg L. Complete sequencing of the p53 gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy. Nat Med. 1995 Oct;1(10):1029–1034. doi: 10.1038/nm1095-1029. [DOI] [PubMed] [Google Scholar]
  7. Borden E. C., Amato D. A., Edmonson J. H., Ritch P. S., Shiraki M. Randomized comparison of doxorubicin and vindesine to doxorubicin for patients with metastatic soft-tissue sarcomas. Cancer. 1990 Sep 1;66(5):862–867. doi: 10.1002/1097-0142(19900901)66:5<862::aid-cncr2820660509>3.0.co;2-r. [DOI] [PubMed] [Google Scholar]
  8. Brodowicz T., Kotz R., Wiltschke C., Zielinski C. C., Ritschl P. Isolated limb perfusion with high-dose tumor necrosis factor-alpha in combination with interferon-gamma and melphalan for nonresectable extremity soft tissue sarcomas: a multicenter trial. J Clin Oncol. 1997 May;15(5):2174–2175. doi: 10.1200/JCO.1997.15.5.2174. [DOI] [PubMed] [Google Scholar]
  9. Canman C. E., Gilmer T. M., Coutts S. B., Kastan M. B. Growth factor modulation of p53-mediated growth arrest versus apoptosis. Genes Dev. 1995 Mar 1;9(5):600–611. doi: 10.1101/gad.9.5.600. [DOI] [PubMed] [Google Scholar]
  10. Crouch G. D., Helman L. J. All-trans-retinoic acid inhibits the growth of human rhabdomyosarcoma cell lines. Cancer Res. 1991 Sep 15;51(18):4882–4887. [PubMed] [Google Scholar]
  11. Delia D., Aiello A., Lombardi L., Pelicci P. G., Grignani F., Grignani F., Formelli F., Menard S., Costa A., Veronesi U. N-(4-hydroxyphenyl)retinamide induces apoptosis of malignant hemopoietic cell lines including those unresponsive to retinoic acid. Cancer Res. 1993 Dec 15;53(24):6036–6041. [PubMed] [Google Scholar]
  12. Di Leonardo A., Linke S. P., Clarkin K., Wahl G. M. DNA damage triggers a prolonged p53-dependent G1 arrest and long-term induction of Cip1 in normal human fibroblasts. Genes Dev. 1994 Nov 1;8(21):2540–2551. doi: 10.1101/gad.8.21.2540. [DOI] [PubMed] [Google Scholar]
  13. Dmitrovsky E. N-(4-hydroxyphenyl)retinamide activation of a distinct pathway signaling apoptosis. J Natl Cancer Inst. 1997 Aug 20;89(16):1179–1181. doi: 10.1093/jnci/89.16.1179. [DOI] [PubMed] [Google Scholar]
  14. Elledge R. M., Gray R., Mansour E., Yu Y., Clark G. M., Ravdin P., Osborne C. K., Gilchrist K., Davidson N. E., Robert N. Accumulation of p53 protein as a possible predictor of response to adjuvant combination chemotherapy with cyclophosphamide, methotrexate, fluorouracil, and prednisone for breast cancer. J Natl Cancer Inst. 1995 Aug 16;87(16):1254–1256. doi: 10.1093/jnci/87.16.1254. [DOI] [PubMed] [Google Scholar]
  15. Engers R., van Roy F., Heymer T., Ramp U., Moll R., Dienst M., Friebe U., Pohl A., Gabbert H. E., Gerharz C. D. Growth inhibition in clonal subpopulations of a human epithelioid sarcoma cell line by retinoic acid and tumour necrosis factor alpha. Br J Cancer. 1996 Feb;73(4):491–498. doi: 10.1038/bjc.1996.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fisher G. J., Datta S. C., Voorhees J. J. Retinoic acid receptor-gamma in human epidermis preferentially traps all-trans retinoic acid as its ligand rather than 9-cis retinoic acid. J Invest Dermatol. 1998 Mar;110(3):297–300. doi: 10.1046/j.1523-1747.1998.00112.x. [DOI] [PubMed] [Google Scholar]
  17. Gabbert H. E., Gerharz C. D., Biesalski H. K., Engers R., Luley C. Terminal differentiation and growth inhibition of a rat rhabdomyosarcoma cell line (BA-HAN-1C) in vitro after exposure to retinoic acid. Cancer Res. 1988 Sep 15;48(18):5264–5269. [PubMed] [Google Scholar]
  18. Gherlinzoni F., Bacci G., Picci P., Capanna R., Calderoni P., Lorenzi E. G., Bernini M., Emiliani E., Barbieri E., Normand A. A randomized trial for the treatment of high-grade soft-tissue sarcomas of the extremities: preliminary observations. J Clin Oncol. 1986 Apr;4(4):552–558. doi: 10.1200/JCO.1986.4.4.552. [DOI] [PubMed] [Google Scholar]
  19. Giandomenico V., Lancillotti F., Fiorucci G., Percario Z. A., Rivabene R., Malorni W., Affabris E., Romeo G. Retinoic acid and IFN inhibition of cell proliferation is associated with apoptosis in squamous carcinoma cell lines: role of IRF-1 and TGase II-dependent pathways. Cell Growth Differ. 1997 Jan;8(1):91–100. [PubMed] [Google Scholar]
  20. Harant H., Lindley I., Uthman A., Ballaun C., Krupitza G., Grunt T., Huber H., Dittrich C. Regulation of interleukin-8 gene expression by all-trans retinoic acid. Biochem Biophys Res Commun. 1995 May 25;210(3):898–906. doi: 10.1006/bbrc.1995.1742. [DOI] [PubMed] [Google Scholar]
  21. Harvey M., Sands A. T., Weiss R. S., Hegi M. E., Wiseman R. W., Pantazis P., Giovanella B. C., Tainsky M. A., Bradley A., Donehower L. A. In vitro growth characteristics of embryo fibroblasts isolated from p53-deficient mice. Oncogene. 1993 Sep;8(9):2457–2467. [PubMed] [Google Scholar]
  22. Hockenbery D., Nuñez G., Milliman C., Schreiber R. D., Korsmeyer S. J. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990 Nov 22;348(6299):334–336. doi: 10.1038/348334a0. [DOI] [PubMed] [Google Scholar]
  23. Kastan M. B., Onyekwere O., Sidransky D., Vogelstein B., Craig R. W. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 1991 Dec 1;51(23 Pt 1):6304–6311. [PubMed] [Google Scholar]
  24. Kerr J. F., Wyllie A. H., Currie A. R. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972 Aug;26(4):239–257. doi: 10.1038/bjc.1972.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lehman T. A., Bennett W. P., Metcalf R. A., Welsh J. A., Ecker J., Modali R. V., Ullrich S., Romano J. W., Appella E., Testa J. R. p53 mutations, ras mutations, and p53-heat shock 70 protein complexes in human lung carcinoma cell lines. Cancer Res. 1991 Aug 1;51(15):4090–4096. [PubMed] [Google Scholar]
  26. Lotan R. Retinoids and apoptosis: implications for cancer chemoprevention and therapy. J Natl Cancer Inst. 1995 Nov 15;87(22):1655–1657. doi: 10.1093/jnci/87.22.1655. [DOI] [PubMed] [Google Scholar]
  27. Lovat P. E., Irving H., Annicchiarico-Petruzzelli M., Bernassola F., Malcolm A. J., Pearson A. D., Melino G., Redfern C. P. Apoptosis of N-type neuroblastoma cells after differentiation with 9-cis-retinoic acid and subsequent washout. J Natl Cancer Inst. 1997 Mar 19;89(6):446–452. doi: 10.1093/jnci/89.6.446. [DOI] [PubMed] [Google Scholar]
  28. Lutzker S. G., Levine A. J. A functionally inactive p53 protein in teratocarcinoma cells is activated by either DNA damage or cellular differentiation. Nat Med. 1996 Jul;2(7):804–810. doi: 10.1038/nm0796-804. [DOI] [PubMed] [Google Scholar]
  29. Mangelsdorf D. J., Thummel C., Beato M., Herrlich P., Schütz G., Umesono K., Blumberg B., Kastner P., Mark M., Chambon P. The nuclear receptor superfamily: the second decade. Cell. 1995 Dec 15;83(6):835–839. doi: 10.1016/0092-8674(95)90199-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Oridate N., Suzuki S., Higuchi M., Mitchell M. F., Hong W. K., Lotan R. Involvement of reactive oxygen species in N-(4-hydroxyphenyl)retinamide-induced apoptosis in cervical carcinoma cells. J Natl Cancer Inst. 1997 Aug 20;89(16):1191–1198. doi: 10.1093/jnci/89.16.1191. [DOI] [PubMed] [Google Scholar]
  31. Repa J. J., Berg J. A., Kaiser M. E., Hanson K. K., Strugnell S. A., Clagett-Dame M. One-step immunoaffinity purification of recombinant human retinoic acid receptor gamma. Protein Expr Purif. 1997 Apr;9(3):319–330. doi: 10.1006/prep.1996.0696. [DOI] [PubMed] [Google Scholar]
  32. Rosenberg S. A., Tepper J., Glatstein E., Costa J., Baker A., Brennan M., DeMoss E. V., Seipp C., Sindelar W. F., Sugarbaker P. The treatment of soft-tissue sarcomas of the extremities: prospective randomized evaluations of (1) limb-sparing surgery plus radiation therapy compared with amputation and (2) the role of adjuvant chemotherapy. Ann Surg. 1982 Sep;196(3):305–315. doi: 10.1097/00000658-198209000-00009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rosso R., Sertoli M. R., Queirolo P., Sanguineti O., Barzacchi M. C., Mariani G. L., Miglio L., Venturini M., Toma S. An outpatient phase I study of a subcutaneous interleukin-2 and intramuscular alpha-2a-interferon combination in advanced malignancies. Ann Oncol. 1992 Jul;3(7):559–563. doi: 10.1093/oxfordjournals.annonc.a058261. [DOI] [PubMed] [Google Scholar]
  34. Ryan J. J., Danish R., Gottlieb C. A., Clarke M. F. Cell cycle analysis of p53-induced cell death in murine erythroleukemia cells. Mol Cell Biol. 1993 Jan;13(1):711–719. doi: 10.1128/mcb.13.1.711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sarkis A. S., Bajorin D. F., Reuter V. E., Herr H. W., Netto G., Zhang Z. F., Schultz P. K., Cordon-Cardo C., Scher H. I. Prognostic value of p53 nuclear overexpression in patients with invasive bladder cancer treated with neoadjuvant MVAC. J Clin Oncol. 1995 Jun;13(6):1384–1390. doi: 10.1200/JCO.1995.13.6.1384. [DOI] [PubMed] [Google Scholar]
  36. Shao Z. M., Dawson M. I., Li X. S., Rishi A. K., Sheikh M. S., Han Q. X., Ordonez J. V., Shroot B., Fontana J. A. p53 independent G0/G1 arrest and apoptosis induced by a novel retinoid in human breast cancer cells. Oncogene. 1995 Aug 3;11(3):493–504. [PubMed] [Google Scholar]
  37. Slichenmyer W. J., Nelson W. G., Slebos R. J., Kastan M. B. Loss of a p53-associated G1 checkpoint does not decrease cell survival following DNA damage. Cancer Res. 1993 Sep 15;53(18):4164–4168. [PubMed] [Google Scholar]
  38. Stewart N., Hicks G. G., Paraskevas F., Mowat M. Evidence for a second cell cycle block at G2/M by p53. Oncogene. 1995 Jan 5;10(1):109–115. [PubMed] [Google Scholar]
  39. Tallman M. S., Andersen J. W., Schiffer C. A., Appelbaum F. R., Feusner J. H., Ogden A., Shepherd L., Willman C., Bloomfield C. D., Rowe J. M. All-trans-retinoic acid in acute promyelocytic leukemia. N Engl J Med. 1997 Oct 9;337(15):1021–1028. doi: 10.1056/NEJM199710093371501. [DOI] [PubMed] [Google Scholar]
  40. Thompson C. B. Apoptosis in the pathogenesis and treatment of disease. Science. 1995 Mar 10;267(5203):1456–1462. doi: 10.1126/science.7878464. [DOI] [PubMed] [Google Scholar]
  41. Toma S., Isnardi L., Raffo P., Dastoli G., De Francisci E., Riccardi L., Palumbo R., Bollag W. Effects of all-trans-retinoic acid and 13-cis-retinoic acid on breast-cancer cell lines: growth inhibition and apoptosis induction. Int J Cancer. 1997 Mar 4;70(5):619–627. doi: 10.1002/(sici)1097-0215(19970304)70:5<619::aid-ijc21>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  42. van der Leede B. M., van den Brink C. E., van der Saag P. T. Retinoic acid receptor and retinoid X receptor expression in retinoic acid-resistant human tumor cell lines. Mol Carcinog. 1993;8(2):112–122. doi: 10.1002/mc.2940080208. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES