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    57 Comprehensive cell mitosis regulation factor

    57 Comprehensive cell mitosis regulation factor

    Comprehensive cell mitosis regulation factor: a computed index named after the cyclin B–CDK1 mitosis-promoting factor; cell-cycle biology, conventional range and limits of interpretation.

    The comprehensive cell mitosis regulation factor is a computed index of the Kolibri method. It is named after the mitosis-promoting factor (MPF), the cyclin B–CDK1 complex that switches a cell from growth into division. Every dividing cell in the body passes through the same cycle: growth (G1), DNA copying (S), preparation (G2) and mitosis (M). The cycle is driven by proteins called cyclins, which are made and destroyed in a regular rhythm, and it is braked by tumour-suppressor proteins such as p53, p21, p27, p15 and p16. When these controls fail, cells divide without limit, and this is a central feature of cancer. The application expresses the overall state of this regulation as one number in conventional units.

    Cell cycleCyclins and CDKsTumour suppressorsComputed index
    What it reflectsregulation of cell division (model estimate)
    Unitsconventional units of the method
    Conventional range3.5–4.6
    Values to reviewbelow 3.5–3.0 or above 4.6–5.0
    Conventional range of the comprehensive cell mitosis regulation factor in this application: below 3.0 low, 3.0 to 3.5 borderline low, 3.5 to 4.6 conventional range, 4.6 to 5.0 borderline high, above 5.0 high Cell mitosis regulation factor (conventional units) Low review with blood-flow data 3.0 Borderline 3.0–3.5 3.5 Conventional range 3.5–4.6 4.6 Borderline 4.6–5.0 5.0 High review with blood-flow data
    Figure 1. Conventional range of the index in this application (conventional units). Values outside the range are a reason for a doctor to review the other results, not a diagnosis.

    The biology behind the name

    In 1971 Masui and Markert found that cytoplasm taken from a mature frog egg makes an immature egg enter division. They called the active substance the maturation-promoting factor. It later turned out to be the same in yeast, frogs and humans: a cyclin-dependent kinase (CDK) bound to a cyclin (Nurse 1990). The CDK is the catalytic subunit. It adds phosphate groups to hundreds of target proteins. The cyclin is the regulatory subunit: the CDK is inactive without it, and the cyclin decides which targets are phosphorylated.

    Different cyclin–CDK pairs act in successive phases (Figure 2). Cyclin D–CDK4/6 responds to growth signals and phosphorylates the retinoblastoma protein (Rb), which releases the genes needed for DNA replication. Cyclin E–CDK2 commits the cell to S phase. Cyclin A–CDK2 completes replication. Cyclin B–CDK1, the MPF itself, starts mitosis: chromosome condensation, breakdown of the nuclear envelope and assembly of the spindle. MPF is held inactive by the kinase Wee1, which phosphorylates CDK1 on tyrosine 15. It is switched on abruptly when the phosphatase Cdc25 removes that phosphate. Because active MPF also activates Cdc25 and inhibits Wee1, entry into mitosis works like an all-or-none switch.

    The cell cycle as a wheel with phases G1, S, G2 and M, the G1/S and G2/M checkpoints, and the cyclin–CDK complexes that drive each phase together with their inhibitors The cell cycle and its drivers G1 S G2 M G1/S checkpoint G2/M checkpoint Cell cycle ≈ 24 h in dividing human cells in culture Cyclin–CDK complexes Cyclin D–CDK4/6 · G1 Responds to growth factors; phosphorylates Rb Inhibited by INK4 proteins: p16, p15 Cyclin E–CDK2 · G1/S Commits the cell to DNA replication Inhibited by CIP/KIP proteins: p21, p27 Cyclin A–CDK2 · S–G2 Completes replication, prepares mitosis Inhibited by p21, p27 Cyclin B–CDK1 (MPF) · G2/M Triggers mitosis; destroyed at anaphase Held off by Wee1, switched on by Cdc25 DNA damage → p53 → p21 → the cycle stops for repair
    Figure 2. The cell cycle. Each phase is driven by its own cyclin–CDK complex, and each complex has its own inhibitors. Checkpoints at the G1/S and G2/M transitions stop the cycle if DNA is damaged or incompletely copied.

    Cyclin destruction: a biochemical oscillator

    Cells break down proteins in two main ways. Long-lived proteins and whole organelles are digested in lysosomes, acidic compartments (pH about 4.5–5) filled with hydrolytic enzymes. Short-lived regulatory proteins, including cyclins, are destroyed in the near-neutral cytosol (pH about 7.2) by the ubiquitin–proteasome system. Ubiquitin is a small protein of 76 amino acids (about 8.6 kDa). A chain of ubiquitin molecules attached to a protein marks it for destruction by the proteasome.

    Cyclin B carries a short sequence of about nine amino acids near its N-terminus, the destruction box. At the transition from metaphase to anaphase, an enzyme complex called the anaphase-promoting complex/cyclosome (APC/C), together with its activator Cdc20, recognises this box and attaches ubiquitin. Cyclin B is then destroyed within minutes, MPF activity collapses, and the cell can separate its chromosomes and divide (Glotzer et al. 1991). Synthesis followed by abrupt destruction makes cyclin levels rise and fall once per cycle (Figure 3). This is why the cell-cycle control system is described as a biochemical oscillator.

    Levels of cyclins D, E, A and B across the phases G1, S, G2 and M: cyclin E peaks at the G1/S transition, cyclin A in G2, cyclin B in mitosis, after which it is abruptly destroyed by APC/C, ubiquitin and the proteasome Cyclin levels across the cell cycle Level G1 S G2 M Cyclin D Cyclin E Cyclin A Cyclin B APC/C + ubiquitin → proteasome
    Figure 3. Schematic levels of the main cyclins during one cycle. Cyclin D (dashed) stays raised for as long as growth signals persist. The other cyclins are synthesised in their own phase and then degraded; the sudden destruction of cyclin B ends mitosis.

    The brakes: p53 and the CDK inhibitors

    Two families of proteins inhibit cyclin–CDK complexes (Sherr & Roberts 1999). They are regulators of the cyclins, not parts of them.

    • CIP/KIP family (p21, p27, p57). The tumour suppressor p53, often called the guardian of the genome, accumulates after DNA damage. It switches on the gene for p21, which blocks CDK2 and CDK1 and stops the cycle until the damage is repaired or the cell is eliminated (el-Deiry et al. 1993). p21 also rises in ageing (senescent) cells. p27 links contact inhibition and transforming growth factor-β (TGF-β) to arrest of the cycle: cells that touch their neighbours stop dividing (Polyak et al. 1994).
    • INK4 family (p16, p15, p18, p19). These proteins bind CDK4 and CDK6 and prevent them from pairing with cyclin D. p15 is induced by TGF-β and helps mediate its growth-inhibiting effect (Hannon & Beach 1994). p16 is one of the genes most frequently lost in human cancers.

    Heat-shock proteins also take part. The chaperone Hsp90, together with its partner Cdc37, binds and stabilises CDK4 (Stepanova et al. 1996). Sustained proliferative signalling and evasion of growth suppressors are two of the recognised hallmarks of cancer (Hanahan & Weinberg 2011). The role of CDKs in cancer is reviewed by Malumbres & Barbacid (2009). Drugs that inhibit CDK4/6 (palbociclib, ribociclib, abemaciclib) are now a standard part of the treatment of hormone-receptor-positive advanced breast cancer.

    How cell proliferation is measured directly

    No blood test measures MPF activity. The reference methods are invasive: they need a sample of tissue obtained by biopsy or surgery. The pathologist counts mitotic figures under the microscope, a key element of tumour grading. Staining for the Ki-67 protein, which is present only in cycling cells, gives the proportion of proliferating cells and is used, for example, in breast cancer (Dowsett et al. 2011). Flow cytometry of DNA content shows the fraction of cells in S phase. The Kolibri index is an estimate obtained from the method's physiological model. It is not a laboratory measurement of these markers.

    Interpretation in this application

    Low values (below 3.5, especially below 3.0)

    The authors of the method regard low values as a signal for the doctor to check for a possible neoplasm. They advise looking at the internal blood-flow parameters (myocardial, muscular, cerebral, hepatoportal, renal, skin and other organs). The organ system involved most often shows reduced blood flow.

    High values (above 4.6, especially above 5.0)

    According to the method's authors, markedly raised values combined with abnormal internal blood-flow parameters may accompany advanced oncological disease. In that situation there are usually obvious symptoms and signs, so history and examination clarify the picture.

    Limits of the index

    This index is not a validated tumour marker. Its relationship with cancer has not been confirmed in published clinical studies. It can neither diagnose nor exclude cancer. A value outside the range should be repeated and reviewed by a doctor together with symptoms and other tests. A normal value is not a reason to skip recommended cancer screening.

    When to see a doctor

    Unexplained weight loss, a new or growing lump, unusual bleeding, a cough or hoarseness lasting more than three weeks, a persistent change in bowel or bladder habits, difficulty swallowing, night sweats or persistent fatigue need prompt medical assessment, whatever this index shows.

    References

    1. Masui Y, Markert CL. Cytoplasmic control of nuclear behavior during meiotic maturation of frog oocytes. J Exp Zool. 1971;177(2):129–145. doi:10.1002/jez.1401770202
    2. Nurse P. Universal control mechanism regulating onset of M-phase. Nature. 1990;344(6266):503–508. doi:10.1038/344503a0
    3. Glotzer M, Murray AW, Kirschner MW. Cyclin is degraded by the ubiquitin pathway. Nature. 1991;349(6305):132–138. doi:10.1038/349132a0
    4. Sherr CJ, Roberts JM. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev. 1999;13(12):1501–1512. doi:10.1101/gad.13.12.1501
    5. el-Deiry WS, Tokino T, Velculescu VE, et al. WAF1, a potential mediator of p53 tumor suppression. Cell. 1993;75(4):817–825. doi:10.1016/0092-8674(93)90500-P
    6. Polyak K, Kato JY, Solomon MJ, et al. p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev. 1994;8(1):9–22. doi:10.1101/gad.8.1.9
    7. Hannon GJ, Beach D. p15INK4B is a potential effector of TGF-β-induced cell cycle arrest. Nature. 1994;371(6494):257–261. doi:10.1038/371257a0
    8. Stepanova L, Leng X, Parker SB, Harper JW. Mammalian p50Cdc37 is a protein kinase-targeting subunit of Hsp90 that binds and stabilizes Cdk4. Genes Dev. 1996;10(12):1491–1502. doi:10.1101/gad.10.12.1491
    9. Malumbres M, Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009;9(3):153–166. doi:10.1038/nrc2602
    10. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–674. doi:10.1016/j.cell.2011.02.013
    11. Dowsett M, Nielsen TO, A'Hern R, et al. Assessment of Ki67 in breast cancer: recommendations from the International Ki67 in Breast Cancer Working Group. J Natl Cancer Inst. 2011;103(22):1656–1664. doi:10.1093/jnci/djr393

    This article is for general information and does not replace a consultation with a doctor. Measurements must be interpreted by a physician together with symptoms and other tests.

    Published on 29 April 2024