Skip to main content
NCBI home page
Journal of Cancer Research and Clinical Oncology logoLink to Journal of Cancer Research and Clinical Oncology
. 1996 Sep;122(9):533–540. doi: 10.1007/BF01213549

An attempt to enhance chemosensitivity of quiescent cell populations in solid tumors by combined treatment with nicotinamide and carbogen

Shin-ichiro Masunaga 1,✉, Koji Ono 1, Mitsuhiko Akaboshi 2, Ken-ichi Kawai 2, Keizo Akuta 3, Masao Takagaki 1, Minoru Suzuki 1, Yuko Kinashi 1, Mitsuyuki Abe 4
PMCID: PMC12200522  PMID: 8781567

Abstract

cis-Diamminedichloroplatinum(II) (cisplatin) was intraperitoneally injected into mice bearing SCC VII or EMT6/KU tumors after ten administrations of 5-bromo-2′-deoxyuridine (BrdU) to label all the proliferating tumor cells. The tumors were excised 1 h after the cisplatin injection, minced, and trypsinized. The tumor cell suspensions were then incubated with cytochalasin-B (a cytokinesis blocker). The micronucleus frequency was determined, using immunofluorescence staining for BrdU. Cells that were not labeled with BrdU were regarded as quiescent. The micronucleus frequency in the total number of tumor cells was determined in tumors that had not been pretreated with BrdU. To modify the sensitivity to cisplatin, nicotinamide was intraperitoneally injected before the administration of cisplatin or mice were placed in a circulating carbogen (95% O2, 5% CO2) chamber for 30 min after cisplatin administration. In both tumor systems, the micronucleus frequency in quiescent cells was lower than that in the total cells. Nicotinamide pretreatment increased the micronucleus frequency in total and in quiescent cells in both tumor systems, and to a higher extent in total cells. The combination of nicotinamide and carbogen increased the micronucleus frequency more markedly than treatment with either nicotinamide or carbogen alone. In total cells of both tumors, the nicotinamide injection increased the uptake of [195mPt]cisplatin. The combined treatment raised the uptake more markedly than did treatment with either agent alone. In total cells of the SCC VII tumor, these increases in micronucleus frequency and the [195mPt]cisplatin uptake following nicotinamide or combined pretreatment were significant. In both tumors, carbogen breathing also elevated the micronucleus frequency to some degree in total and quiescent cells and the [195mPt]cisplatin uptake in total cells. The combined nicotinamide and carbogen treatment was considered to be useful for sensitizing tumor cells to chemotherapy with cisplatin in vivo.

Key words: Quiescent cell, cis-Diamminedichloroplatinum(II), Nicotinamide, Carbogen, Radiolabeled cisplatin

Abbreviations

Q

cells quiescent cells

P

cells proliferating cells

References

  1. Akaboshi M, Kawai K, Maki H, Akuta K, Ujeno Y, Miyahara T (1992) The number of platinum atoms binding to DNA, RNA and protein molecules of Hela cells treated with cisplatin at its mean lethal concentration. Jpn J Cancer Res 83:522–526 [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown JM (1979) Evidence for acutely hypoxic cells in mouse tumours, and a possible mechanism of reoxygenation. Br J Radiol 52:650–656 [DOI] [PubMed] [Google Scholar]
  3. Bush RS, Jenkins RDT, Allt WEC, Beal FA, Bean HA, Dembo AJ, Pringle JF (1978) Definitive evidence for hypoxic cells influencing cure in cancer therapy. Br J Cancer 37 [Suppl III]: 302–306 [PMC free article] [PubMed] [Google Scholar]
  4. Chaplin DJ, Horsman MR, Trotter MJ (1990) Effect of nicotinamide on the microregional heterogeneity of oxygen delivery within a murine tumor. J Natl Cancer Inst 82:672–676 [DOI] [PubMed] [Google Scholar]
  5. Coleman CN (1988) Review. Hypoxia in tumors: a paradigm for the approach to biochemical and physiologic heterogeneity. J Natl Cancer Inst 80:310–317 [DOI] [PubMed] [Google Scholar]
  6. Dethlefsen LA (1980) In quest of the quaint quiescent cells. In: Meyn RE, Withers HR (eds) Radiation biology in cancer research. Raven, New York, pp 415–435 [Google Scholar]
  7. Fenech M, Morley A (1985) Solutions to the kinetic problem in the micronucleus assay. Cytobios 43:233–246 [PubMed] [Google Scholar]
  8. Fenton BM (1995) The effects of carbogen and nicotinamide on intravascular oxyhaemoglobin saturations in SCC VII and KHT murine tumours. Br J Cancer 71:945–949 [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Freyer JP (1992) Spheroids in radiobiology research. In: Bjerkvik R (ed) Spheroid culture in cancer research. CRC, London, pp 217–275 [Google Scholar]
  10. Henk JM, Smith CW (1977) Radiotherapy and hyperbaric oxygen in head and neck cancer. Lancet 2:104–105 [DOI] [PubMed] [Google Scholar]
  11. Hermens AF, Barendsen GW (1978) The proliferative status and clonogenic capacity of tumor cells in a transplantable rhabdomyosarcoma of the rat before and after irradiation with 800 rad of X-rays. Cell Tissue Kinet 11: 83–100 [DOI] [PubMed] [Google Scholar]
  12. Horsman MR, Wood PJ, Chaplin DJ, Brown JM, Overgaard J (1990) The potentiation of radiation damage by nicotinamide in the SCCVII tumour in vivo. Radiother Oncol 18:49–57 [DOI] [PubMed] [Google Scholar]
  13. Horsman MR, Nordsmark M, Khalil AA, Hill SA, Chaplin DJ, Siemann DW, Overgaard J (1994) Reducing acute and chronic hypoxia in tumours by combining nicotinamide with carbogen breathing. Acta Oncol 33:371–376 [DOI] [PubMed] [Google Scholar]
  14. Jackson RC (1989) The problem of the quiescent cancer cell. Adv Enzyme Regul 29:27–46 [DOI] [PubMed] [Google Scholar]
  15. Jain RK (1989) Delivery of novel therapeutic agents in tumors: physiological barriers and strategies. J Natl Cancer Inst 81:570–576 [DOI] [PubMed] [Google Scholar]
  16. Kallinowski F, Schlenger KH, Runkel S, Kloes S, Stohrer M, Okunnieff P, Vaupel P (1989) Blood flow, metabolism, cellular microenvironment and growth rate of human tumour xenografts. Cancer Res 49:3759–3764 [PubMed] [Google Scholar]
  17. Kallman RF, Comb CA, Franco AJ, Furlong BM, Kelly SD, Kemper HL, Miller RG, Rapachietta D, Schoenfeld D, Takahashi M (1980) Evidence for the recruitment of noncycling clonogenic tumor cells. In: Meyn RE, Withers HR (eds) Radiation biology in cancer research. Raven, New York, pp 397–414 [Google Scholar]
  18. Kawai K, Maki H, Ehrlich W, Akaboshi M (1989) Synthesis of195mPt radiolabeledcis-diamminedichloroplatinum(II) of high chemical and radiochemical purity using high performance liquid chromatography. J Radioanal Nucl Chem Lett 136:67–74 [Google Scholar]
  19. Kjellen E, Joiner MC, Collier JM, Johns H, Rojas AA (1991) A therapeutic benefit from combining normobaric carbogen or oxygen with nicotinamide in fractionated X-ray treatments. Radiother Oncol 22:81–91 [DOI] [PubMed] [Google Scholar]
  20. Lajtha LG (1963) On the concepts of the cell cycle. Cell Comp Physiol 62:143–145 [PubMed] [Google Scholar]
  21. Little JB (1977) Factors influencing the repair of potentially lethal radiation damage in growth-inhibited human cells. Radiat Res 70:705–706 [PubMed] [Google Scholar]
  22. Luk CK, Keng PC, Sutherland RM (1985) Regrowth and radiation sensitivity of quiescent cells isolated from EMT6/Ro-fed plateau monolayers. Cancer Res 45:1020–1025 [PubMed] [Google Scholar]
  23. Masunaga S, Ono K, Abe M (1991) A method for selective measurement of the radiosensitivity of quiescent cells in solid tumors—combination of immunofluorescence staining to BrdU and micronucleus assay. Radiat Res 125:243–247 [PubMed] [Google Scholar]
  24. Masunaga S, Ono K, Abe M (1993) The detection and modification of the hypoxic fraction in quiescent cell populations in murine solid tumours. Br J Radiol 66:918–926 [DOI] [PubMed] [Google Scholar]
  25. Ono K, Wandl EO, Tsutsui K, Sasai K, Abe M (1989) The correlation between cell survival curve and dose response curve of micronucleus (MN) frequency. Strahlenther Onkol 165:824–827 [PubMed] [Google Scholar]
  26. Overgaard J (1989) Sensitization of hypoxic tumor cells—clinical experience. Int J Radiat Biol 56:801–811 [DOI] [PubMed] [Google Scholar]
  27. Shibamoto Y, Yukawa K, Tsutsui K, Takahashi M, Abe M (1986) Variation in the hypoxic fraction among mouse tumors of different types, size, and sites. Jpn J Cancer Res 77:908–915 [PubMed] [Google Scholar]
  28. Steel GG (1977) Cell population kinetics of tumors in experimental animals. In: Steel GG (eds) Growth kinetics of tumours, Clarendon, Oxford, pp 146–216 [Google Scholar]
  29. Wallen CA, Higashikubo R, Dethlefsen LA (1984) Murine mammary cells in vitro. II. Recruitment of quiescent cells. Cell Tissue Kinet 17:79–89 [DOI] [PubMed] [Google Scholar]
  30. Wallen CA, Ridinger DN, Dethlefsen LA (1985) Heterogeneity of X-ray cytotoxicity in proliferating and quiescent murine mammary carcinoma cells. Cancer Res 45:3064–3069 [PubMed] [Google Scholar]
  31. Wilson KM, Keng PC (1989) Radiation-induced DNA damage and repair in quiescent and proliferating human tumor cells in vitro. Int J Radiat Biol 55:385–395 [DOI] [PubMed] [Google Scholar]

Articles from Journal of Cancer Research and Clinical Oncology are provided here courtesy of Springer

RESOURCES