Tumor-Specific S/G2-Phase Cell Cycle Arrest of Cancer Cells by Methionine Restriction

  • Robert M. HoffmanEmail author
  • Shuya Yano
Part of the Methods in Molecular Biology book series (MIMB, volume 1866)


Cancer cells require elevated amounts of methionine (MET) and arrest their growth under conditions of MET restriction (MR). This phenomenon is termed MET dependence. Fluorescence-activated cell sorting (FACS) first indicated that the MET-dependent SV40-transformed cancer cells were arrested in the S and G2 phases of the cell cycle when under MR. This is in contrast to a G1-phase accumulation of cells, which occurs only in MET-supplemented medium at very high cell densities and which is similar to the G1 cell-cycle block which occurs in cultures of normal fibroblasts at high density. When the human PC-3 prostate carcinoma cell line was cultured in MET-free, homocysteine-containing (METHCY+) medium, there was an extreme increment in DNA content without cell division indicating that the cells were blocked in S phase. Recombinant methioninase (rMETase) treatment of cancer cells also selectively trapped cancer cells in S/G2: The cell cycle phase of the cancer cells was visualized with the fluorescence ubiquitination cell cycle indicator (FUCCI). At the time of rMETase-induced S/G2-phase trap, identified by the cancer cells’ green fluorescence by FUCCI imaging, the cancer cells were administered S-phase-dependent chemotherapy drugs, which interact with DNA or block DNA synthesis such as doxorubicin, cisplatin, or 5-fluorouracil (5-FU) and which were highly effective in killing the cancer cells. In contrast, treatment of cancer cells with drugs in the presence of MET, only led to the majority of the cancer cell population being blocked in G0/G1 phase, identified by the cancer cells becoming red fluorescent in the FUCCI system. The G0/G1 blocked cells were resistant to the chemotherapy. MR has the potential for highly effective cell-cycle-based treatment strategy for cancer in the clinic.

Key words

Cancer cells Methionine dependence Methionine restriction Cell cycle Arrest S/G2 phase, chemotherapy 


  1. 1.
    Hoffman RM (2015) Development of recombinant methioninase to target the general cancer-specific metabolic defect of methionine dependence: a 40-year odyssey. Expert Opin Biol Ther 15:21–31CrossRefGoogle Scholar
  2. 2.
    Kamely D, Littlefield JW, Erbe R (1973) Regulation of 5-methyltetrahydrofolate: homocysteine methyltransferase activity by methionine, vitamin B12, and folate in cultured baby hamster kidney cells. Proc Natl Acad Sci U S A 70:2585–2589CrossRefGoogle Scholar
  3. 3.
    Hoffman RM, Jacobsen SJ (1980) Reversible growth arrest in simian virus 40-transformed human fibroblasts. Proc Natl Acad Sci U S A 77:7306–7310CrossRefGoogle Scholar
  4. 4.
    Guo HY, Herrera H, Hoffman RM (1993) Unchecked DNA synthesis and blocked cell division induced by methionine deprivation in a human prostate cancer cell line. In Vitro Cell Dev Biol 29A:359–361CrossRefGoogle Scholar
  5. 5.
    Guo H, Lishko V, Herrera H, Groce A, Kubota T, Hoffman RM (1993) Therapeutic tumor-specific cell-cycle block induced by methionine starvation in vivo. Cancer Res 53:5676–5679PubMedGoogle Scholar
  6. 6.
    Stern PH, Hoffman RM (1986) Enhanced in vitro selective toxicity of chemotherapeutic agents for human cancer cells based on a metabolic defect. J Natl Cancer Inst 76:629–639CrossRefGoogle Scholar
  7. 7.
    Yano S, Li S, Han Q, Tan Y, Bouvet M, Fujiwara T, Hoffman RM (2014) Selective methioninase-induced trap of cancer cells in S/G2 phase visualized by FUCCI imaging confers chemosensitivity. Oncotarget 5:8729–8736CrossRefGoogle Scholar
  8. 8.
    Hoffman RM, Jacobsen SJ, Erbe RW (1979) Reversion to methionine independence in simian virus 40-transformed human and malignant rat fibroblasts is associated with altered ploidy and altered properties of transformation. Proc Natl Acad Sci U S A 76:1313–1317CrossRefGoogle Scholar
  9. 9.
    Kraemer PM, Deaven LL, Crissman HA, Van Dilla MA (1972) In: Du-Praw EJ (ed) Advances in cell and molecular biology, vol 2. Academic Press, New York, pp 47–108Google Scholar
  10. 10.
    Guo HY, Herrera H, Groce A, Hoffman RM (1993) Expression of the biochemical defect of methionine dependence in fresh patient tumors in primary histoculture. Cancer Res 53:2479–2483PubMedGoogle Scholar
  11. 11.
    Yano S, Zhang Y, Miwa S, Tome Y, Hiroshima Y, Uehara F, Yamamoto M, Suetsugu A, Kishimoto H, Tazawa H, Zhao M, Bouvet M, Fujiwara T, Hoffman RM (2014) Spatial-temporal FUCCI imaging of each cell in a tumor demonstrates locational dependence of cell cycle dynamics and chemoresponsiveness. Cell Cycle 13:2110–2119CrossRefGoogle Scholar
  12. 12.
    Yano S, Miwa S, Mii S, Hiroshima Y, Uehara F, Yamamoto M, Kishimoto H, Tazawa H, Bouvet M, Fujiwara T, Hoffman RM (2014) Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging. Cell Cycle 13:953–960CrossRefGoogle Scholar
  13. 13.
    Yano S, Tazawa H, Hashimoto Y, Shirakawa Y, Kuroda S, Nishizaki M, Kishimoto H, Uno F, Nagasaka T, Urata Y, Kagawa S, Hoffman RM, Fujiwara T (2013) A genetically engineered oncolytic adenovirus decoys and lethally traps quiescent cancer stem-like cells into S/G2/M phases. Clin Cancer Res 19:6495–6505CrossRefGoogle Scholar
  14. 14.
    Yano S, Takehara K, Zhao M, Tan Y, Han Q, Li S, Bouvet M, Fujiwara T, Hoffman RM (2016) Tumor-specific cell-cycle decoy by Salmonella typhimurium A1-R combined with tumor-selective cell-cycle trap by methioninase overcome tumor intrinsic chemoresistance as visualized by FUCCI imaging. Cell Cycle 15:1715–1723CrossRefGoogle Scholar
  15. 15.
    Castedo M, Perfettini JL, Roumier T, Andreau K, Medema R, Kroemer G (2004) Cell death by mitotic catastrophe: a molecular definition. Oncogene 23:2825–2837CrossRefGoogle Scholar
  16. 16.
    Chow JPH, Poon RYC (2010) Mitotic catastrophe. In: Enders G (ed) Cell cycle deregulation in cancer. Springer, New York, pp 79–96CrossRefGoogle Scholar
  17. 17.
    Vakifahmetoglu H, Olsson M, Zhivotovsky B (2008) Death through a tragedy: mitotic catastrophe. Cell Death Differ 15:1153–1162CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.AntiCancer, Inc.San DiegoUSA
  2. 2.Department of SurgeryUniversity of CaliforniaSan DiegoUSA

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