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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1991 Jan;82(1):118–126. doi: 10.1111/j.1349-7006.1991.tb01754.x

Mechanism of the Combined Antitumor Effect of Natural Human Tumor Necrosis Factor‐α and Natural Human Interferon‐α on Cell Cycle Progression

Masahiko Muro 1, Yoshio Naomoto 1, Kunzo Orita 1
PMCID: PMC5918212  PMID: 1825650

Abstract

We have studied the mechanism of the synergistic effect of the combination of tumor necrosis factor‐α (TNF‐α) and interferon‐a (IFN‐α) on cell cycle progression using two‐parameter flow cytometry in vitro and an immunohistochemical staining method in vivo. The cells used were human colon cancer cell line RPMI 4788 in vitro and in vivo, and human breast cancer cell line MX‐1 and human renal cancer cell line NAMKO‐1 in vivo. In the in vitro experiment, the cell cycle progressed normally as time elapsed in the control group. However, in the group treated with TNF‐α and IFN‐α in combination (combination group), it appeared that the transition from the S phase to the G2/M phase was blocked, and the cells that accumulated in the S phase died. In the in vivo experiment with male nude mice of a CD‐1 genetic background, the antitumor effect on all three kinds of cancer cells was significantly greater in the combination group than in the control group. The cell labeling index on staining with bromodeoxyuridine in the combination group became markedly larger and the mitotic index smaller than in the other groups. From these results, it was concluded that in the combination group, both in vitro and in vivo, tumor cells markedly accumulated in the S phase and their progression from the S phase to the G2/M phase in the cell cycle was inhibited.

Keywords: Tumor necrosis factor‐α, Interferon‐α, Cell cycle progression

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REFERENCES

  • 1. ) Bevilacqua , M. P. , Pober , J. S. , Majeau , G. R. , Fiers , W. , Cotran , R. S. and Gimbrone , M. A. , Jr.Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endothelium: characterization and comparison with the action of interleukin‐1 . Proc, Natl. Acad. Sci. USA , 83 , 4533 – 4537 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. ) Beutler , B. A. and Cerami , A.Recombinant interleukin 1 suppressed lipoprotein lipase activity in 3T3‐L1 cells . J. Immunol , 135 , 3969 – 3971 ( 1985. ). [PubMed] [Google Scholar]
  • 3. ) Kawakami , M. , Pekala , P. H. , Lane , M. D. and Cerami , A.Lipoprotein lipase suppression in 3T3‐L1 cells by an endotoxin‐induced mediatorfrom exudate cells . Proc. Natl. Acad. Sci. USA , 79 , 912 – 916 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. ) Dayer , J. M. , Beutler , B. and Cerami , A.Cachectin/tumor necrosis factor stimulates collagenase and prostaglandin E2 production by human synovial cells and dermal fibroblasts . J. Exp. Med. , 162 , 2163 – 2168 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. ) Mizel , S. B. , Dayer , J. M. , Krane , S. M. and Mergenhagen , S. E.Stimulation of rheumatoid synovial cell collagenase prostaglandin production by partially purified lymphocyte activating factor (interleukin‐1) . Proc. Natl. Acad. Sci. USA , 78 , 2474 – 2478 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Dinarello , C. A. , Cannon , J. G. , Wolff , S. M. , Bernheim , H. A. , Beutler , B. , Cerami , A. , Figari , I. S. , Palladino , M. A. and Oapos;Connor , J. V.Tumor necrosis factor (cachectin) is an endogenous pyrogen and induces production of interleukin‐1 . J. Exp. Med. , 163 , 1433 – 1450 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. ) Gowen , M. , Wood , D. D. , Ihrie , E. J. , McGuire , M. K. B. and Russell , R. G. G.An IL‐1 like factor stimulates bone resorption in vitro . Nature , 306 , 378 – 380 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Bertolini , D. R. , Nedwh , G. E. , Bringman , T. S. , Smith , D. D. and Mundy , G. R.Stimulation of bone resorption and inhibition of bone formation in vitro by human tumor necrosis factor . Nature , 319 , 516 – 518 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Shalaby , M. R. , Aggarwal , B. B. , Rinderknicht , E. , Svedersky , L. P.Finkle , B. S. and Palladino , M. A. , Jr.Activation of human polymorphonuclear neutrophil functions by interferon‐7 and tumor necrosis factor . J, Immunol , 135 , 2069 – 2073 ( 1985. ). [PubMed] [Google Scholar]
  • 10. ) Sugarman , B. J. , Aggarwal , B. B. , Hass , P. E. , Figari , I. S. , Palladino , M. A. , Jr. and Shepard , H. M.Recombinant human tumor necrosis factor‐α: effects on proliferation of normal and transformed cells in vitro . Science , 230 , 943 – 945 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Vilcék , J. , Palombella , V. J. , Henriksen‐DeStefano , D. , Swenson , C. , Feinman , R. , Hirai , M. and Tsujimoto , M.Fibroblast growth enhancing activity of tumor necrosis factor and its relationship to other polypeptide growth factors . J. Exp. Med. , 163 , 632 – 643 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. ) Williamson , B. D. , Carswell , E. A. , Rubin , B. Y. , Prendergast , J. S. and Old , L. J.Human tumor necrosis factor produced by human B‐cell lines: synergistic cytotoxic interaction with human interferon , Proc. Natl. Acad. Sci. USA , 80 , 5397 – 5401 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. ) Balkwil , F. R. , Lee , A. , Aldam , G. , Moodie , E. , Thomas , J. A. , Tavernier , J. and Fiers , W.Human tumor xenografts treated with recombinant human tumor necrosis factor alone or in combination with interferon . Cancer Res. , 46 , 3990 – 3993 ( 1986. ). [PubMed] [Google Scholar]
  • 14. ) Naomoto , Y. , Tanaka , N. , Fuchimoto , S. and Orita , K.In vitro synergistic effects of natural human tumor necrosis factor and natural human interferon‐α . Jpn, J. Cancer Res. , 78 , 87 – 92 ( 1987. ). [PubMed] [Google Scholar]
  • 15. ) Orita , K. , Ando , S. and Kurimoto , M.Synergism between human tumor necrosis factor and human interferon‐α: effects on cells in culture . Acta Med. Okayama , 41 , 155 – 160 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) McIntosh , J. K. , Mule , J.J. , Krosnick , J. A. and Rosenberg , S. A.Combination cytokine immunotherapy with tumor necrosis factor‐α, interleukin 2, andα‐interferon and its synergistic antitumor effects in mice Cancer Res. , 49 , 1408 – 1414 ( 1989. ). [PubMed] [Google Scholar]
  • 17. ) Naomoto , Y. , Tanaka , N. and Orita , K.Antitumor effect of natural human tumor necrosis factor‐α and natural human interferon‐α in combination against human cancer transplanted into nude mice . Acta Med. Okayama , 43 , 211 – 221 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Fuchimoto , S. , Itano , S. , Kimura , T. , Iwagaki , H. , Maeda , T. , Hamada , H. , Kurimoto , M. and Orita , K.Phase 1 and early phase 2 clinical study of a new antitumor lymphokine produced by stimulated B cell line, BALL‐1 . Proc. Am. Soc. Clin. Oncol , 22 , 910 ( 1986. ). [Google Scholar]
  • 19. ) Moore , G. E. and Koike , E.Growth of human tumor cells in vitro and in vivo . Cancer , 17 , 11 – 20 ( 1964. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Ovejera , A. A. , Houchens , D. P. and Barker , A. D.Chemotherapy of human tumor xenografts in genetically athymic mice . Ann. Clin. Lab. Sci. , 8 , 50 – 56 ( 1978. ). [PubMed] [Google Scholar]
  • 21. ) Fukuda , S. , Ando , S. , Sanou , O. , Taniai , M. , Fujii , M. , Masaki , N. , Nakamura , K. , Ando , O. , Torigoe , K. , Sugimoto , T. and Kurimoto , M.Simultaneous production of natural human tumor necrosis factor‐α, ‐β and interferon‐α from BALL‐ 1 cells stimulated by HVJ . Lymphokine Res. , 7 , 175 – 185 ( 1988. ). [PubMed] [Google Scholar]
  • 22. ) Fukuda , S. , Yamane , Y. , Kouguchi , M. , Ando , S. , Sato , Y. and Kurimoto , M.Purification and characterization of natural human tumor necrosis factor from HVJ stimulated BALL‐1 cells . Chem. Express , 2 , 101 – 104 ( 1987. ). [Google Scholar]
  • 23. ) Nobuhara , M. , Kanamori , T. , Nagase , Y. , Nii , A. , Morishita , H. , Toyama , J. , Ando , S. and Kurimoto , M.The expression of human tumor necrosis factor in E . coli. Nucleic Acid Symp. Ser. , 17 , 131 – 134 ( 1986. ). [PubMed] [Google Scholar]
  • 24. ) Sohmura , Y. , Nakano , K. and Abe , S.Antitumor activity of recombinant human tumor necrosis factor . Jpn. J. Cancer Chemother. , 12 , 160 – 162 ( 1985. ). [PubMed] [Google Scholar]
  • 25. ) Lee , S. H. , Aggarwell , B. B. , Rinderknecht , E. , Assisi , F. and Chiu , H.The synergistic anti‐proliferative effect of γ‐interferon and human lymphotoxin . J. Immunol. , 133 , 1083 – 1090 ( 1984. ). [PubMed] [Google Scholar]
  • 26. ) Dean , P. N. , Gray , J. W. and Dolbeare , F. A.The analysis and interpretation of DNA distributions measured by flow cytometry . Cytometry , 3 , 188 – 198 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Naomoto , Y. , Kondo , H. , Tanaka , N. and Orita , K.Novel experimental models of human cancer metastasis in nude mice; lung metastasis, intraabdominal carcinomatosis with ascites and liver metastasis . J. Cancer Res., Clin. Oncol , 113 , 544 – 549 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Sugarman , B. J. , Aggarwal , B. B. , Hass , P. E. , Figari , I. S. , Palladino , M. A. , Jr. and Shepard , H. M.Recombinant human tumor necrosis factor‐α: effects on proliferation of normal and transformed cells in vitro . Science , 230 , 943 – 945 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Nakano , K. , Abe , S. and Sohmura , Y.Recombinant human tumor necrosis factor‐I. Cytotoxic activity in vitro . Int. J. Immunopharmacol. , 8 , 347 – 355 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Sohmura , Y. , Nakata , K. , Yoshida , H. , Kashimoto , S. , Matsui , Y. and Furuichi , H.Recombinant human tumor necrosis factor‐II. Antitumor effect on murine and human tumors transplanted in mice . Int. J. Immunopharmacol. , 8 , 357 – 368 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Manda , T. , Shimomura , K. , Mukumoto , S. , Kobayashi , K. , Mizuta , T. , Hirai , O. , Matsumoto , S. , Oku , T. , Nishigaki , F. , Mori , J. and Kikuchi , H.Recombinant human tumor necrosis factor‐α: evidence of an indirect mode of antitumor activity . Cancer Res. , 47 , 3707 – 3711 ( 1987. ). [PubMed] [Google Scholar]
  • 32. ) Shimomura , K. , Manda , T. , Mukumoto , S. , Kobayashi , K. , Nakano , K. and Mori , J.Recombinant human tumor necrosis factor‐α: thrombus formation is a cause of antitumor activity . Int. J. Cancer , 41 , 243 – 247 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Fiers , W. , Brouckaert , P. , Devos , R. , Fransen , L. , Leroux‐Roels , G. , Remaunt , E. , Suffys , P. , Tavernier , J. , Van der Heyden , J. and Van Roy , F.Lymphokines and monokines in anticancer therapy . Cold Spring Harbor Symp. Quant. Biol , 51 , 589 – 595 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Aggarwal , B. B. , Eessalu , T. E. and Hass , P. E.Characterization of receptors for human tumor necrosis factor and their regulation by γ‐interferon . Nature , 318 , 665 – 667 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 35. ) Ruggiero , V. , Tavernier , J. , Fiers , W. and Baglioni , C.Induction of the synthesis of tumor necrosis factor receptors by interferon‐γ . J. Immunol. , 136 , 2445 – 2450 ( 1986. ). [PubMed] [Google Scholar]
  • 36. ) Tsujimoto , M.Yip , Y. K. and Vilcék , J.Interferon‐enhances expression of cellular receptors for tumor necrosis factor . J. Immunol. , 136 , 2441 – 2444 ( 1986. ). [PubMed] [Google Scholar]
  • 37. ) Ruff , M. R. and Gifford , G. E.Tumor necrosis factor . Lymphokines , 2 , 235 – 272 ( 1981. ). [Google Scholar]
  • 38. ) Darzykiewics , Z. , Williamson , B. , Carswell , E. A. and Old , L. J.Cell cycle‐specific effects of tumor necrosis factor . Cancer Res. , 44 , 83 – 90 ( 1984. ). [PubMed] [Google Scholar]
  • 39. ) Creasey , A. A. , Bartholomew , J. C. and Merigan , T. C.Role of G0–G1 arrest in the inhibition of tumor cell growth by interferon . Proc. Natl. Acad. Sci. USA , 77 , 1471 – 1475 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40. ) Beutler , B. , Groenwald , D. , Holmes , J. D. , Chang , M. , Pan , Y. C. E. , Mathison , J. , Ulevitch , R. and Cerami , A.Identity of tumor necrosis factor and the macrophage‐secreted factor cachectin . Nature , 316 , 552 – 553 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 41. ) Beutler , B. and Cerami , A.Cachectin and tumor necrosis factor as two sides of the same biological coin . Nature , 320 , 584 – 588 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 42. ) Beutler , B. , Milsark , I. W. and Cerami , A.Cachectin/tumor necrosis factor: Production, distribution and metabolic fate in vivo . J. Immunol. , 135 , 3972 – 3977 ( 1985. ). [PubMed] [Google Scholar]

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