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J Cell Biol. Nov 1, 1993; 123(3): 513–522.
PMCID: PMC2200129

The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins

Abstract

The antigen defined by mAb Ki-67 is a human nuclear protein the expression of which is strictly associated with cell proliferation and which is widely used in routine pathology as a "proliferation marker" to measure the growth fraction of cells in human tumors. Ki-67 detects a double band with apparent molecular weights of 395 and 345 kD in immunoblots of proteins from proliferating cells. We cloned and sequenced the full length cDNA, identified two differentially spliced isoforms of mRNA with open reading frames of 9,768 and 8,688 bp encoding for this cell proliferation-associated protein with calculated molecular weights of 358,761 D and 319,508 D, respectively. New mAbs against a bacterially expressed part and a synthetic polypeptide deduced from the isolated cDNA react with the native Ki-67 antigen, thus providing a circle of evidence that we have cloned the authentic Ki-67 antigen cDNA. The central part of the Ki-67 antigen cDNA contains a large 6,845-bp exon with 16 tandemly repeated 366-bp elements, the "Ki-67 repeats", each including a highly conserved new motif of 66 bp, the "Ki-67 motif", which encodes for the epitope detected by Ki-67. Computer analysis of the nucleic acid and the deduced amino acid sequence of the Ki-67 antigen confirmed that the cDNA encodes for a nuclear and short-lived protein without any significant homology to known sequences. Ki-67 antigen-specific antisense oligonucleotides inhibit the proliferation of IM-9 cell line cells, indicating that the Ki-67 antigen may be an absolute requirement for maintaining cell proliferation. We conclude that the Ki-67 antigen defines a new category of cell cycle-associated nuclear nonhistone proteins.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ball TK, Suh Y, Benedik MJ. Disulfide bonds are required for Serratia marcescens nuclease activity. Nucleic Acids Res. 1992 Oct 11;20(19):4971–4974. [PMC free article] [PubMed]
  • Bavisotto L, Kaushansky K, Lin N, Hromas R. Antisense oligonucleotides from the stage-specific myeloid zinc finger gene MZF-1 inhibit granulopoiesis in vitro. J Exp Med. 1991 Nov 1;174(5):1097–1101. [PMC free article] [PubMed]
  • Bouzubar N, Walker KJ, Griffiths K, Ellis IO, Elston CW, Robertson JF, Blamey RW, Nicholson RI. Ki67 immunostaining in primary breast cancer: pathological and clinical associations. Br J Cancer. 1989 Jun;59(6):943–947. [PMC free article] [PubMed]
  • Braun N, Papadopoulos T, Müller-Hermelink HK. Cell cycle dependent distribution of the proliferation-associated Ki-67 antigen in human embryonic lung cells. Virchows Arch B Cell Pathol Incl Mol Pathol. 1988;56(1):25–33. [PubMed]
  • Brown DC, Gatter KC. Monoclonal antibody Ki-67: its use in histopathology. Histopathology. 1990 Dec;17(6):489–503. [PubMed]
  • Bruno S, Darzynkiewicz Z. Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells. Cell Prolif. 1992 Jan;25(1):31–40. [PubMed]
  • Celis JE, Madsen P. Increased nuclear cyclin/PCNA antigen staining of non S-phase transformed human amnion cells engaged in nucleotide excision DNA repair. FEBS Lett. 1986 Dec 15;209(2):277–283. [PubMed]
  • Chelsky D, Ralph R, Jonak G. Sequence requirements for synthetic peptide-mediated translocation to the nucleus. Mol Cell Biol. 1989 Jun;9(6):2487–2492. [PMC free article] [PubMed]
  • Chou PY, Fasman GD. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. [PubMed]
  • Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulaziz Z, MacDonald S, Pulford KA, Stein H, Mason DY. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem. 1984 Feb;32(2):219–229. [PubMed]
  • Dingwall C, Laskey RA. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. [PubMed]
  • du Manoir S, Guillaud P, Camus E, Seigneurin D, Brugal G. Ki-67 labeling in postmitotic cells defines different Ki-67 pathways within the 2c compartment. Cytometry. 1991;12(5):455–463. [PubMed]
  • Fontana D, Bellina M, Gubetta L, Fasolis G, Rolle L, Scoffone C, Porpiglia F, Colombo M, Tarabuzzi R, Leonardo E. Monoclonal antibody Ki-67 in the study of the proliferative activity of bladder carcinoma. J Urol. 1992 Oct;148(4):1149–1151. [PubMed]
  • Frohman MA, Dush MK, Martin GR. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. [PMC free article] [PubMed]
  • Gerdes J. Ki-67 and other proliferation markers useful for immunohistological diagnostic and prognostic evaluations in human malignancies. Semin Cancer Biol. 1990 Jun;1(3):199–206. [PubMed]
  • Gerdes J, Schwab U, Lemke H, Stein H. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer. 1983 Jan 15;31(1):13–20. [PubMed]
  • Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H. Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol. 1984 Oct;133(4):1710–1715. [PubMed]
  • Gerdes J, Stein H, Pileri S, Rivano MT, Gobbi M, Ralfkiaer E, Nielsen KM, Pallesen G, Bartels H, Palestro G, et al. Prognostic relevance of tumour-cell growth fraction in malignant non-Hodgkin's lymphomas. Lancet. 1987 Aug 22;2(8556):448–449. [PubMed]
  • Gerdes J, Li L, Schlueter C, Duchrow M, Wohlenberg C, Gerlach C, Stahmer I, Kloth S, Brandt E, Flad HD. Immunobiochemical and molecular biologic characterization of the cell proliferation-associated nuclear antigen that is defined by monoclonal antibody Ki-67. Am J Pathol. 1991 Apr;138(4):867–873. [PMC free article] [PubMed]
  • Grogan TM, Lippman SM, Spier CM, Slymen DJ, Rybski JA, Rangel CS, Richter LC, Miller TP. Independent prognostic significance of a nuclear proliferation antigen in diffuse large cell lymphomas as determined by the monoclonal antibody Ki-67. Blood. 1988 Apr;71(4):1157–1160. [PubMed]
  • Guillaud P, du Manoir S, Seigneurin D. Quantification and topographical description of Ki-67 antibody labelling during the cell cycle of normal fibroblastic (MRC-5) and mammary tumour cell lines (MCF-7). Anal Cell Pathol. 1989 Feb;1(1):25–39. [PubMed]
  • Hall PA, Richards MA, Gregory WM, d'Ardenne AJ, Lister TA, Stansfeld AG. The prognostic value of Ki67 immunostaining in non-Hodgkin's lymphoma. J Pathol. 1988 Mar;154(3):223–235. [PubMed]
  • Hall PA, Levison DA, Woods AL, Yu CC, Kellock DB, Watkins JA, Barnes DM, Gillett CE, Camplejohn R, Dover R, et al. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990 Dec;162(4):285–294. [PubMed]
  • Hall PA, McKee PH, Menage HD, Dover R, Lane DP. High levels of p53 protein in UV-irradiated normal human skin. Oncogene. 1993 Jan;8(1):203–207. [PubMed]
  • Harel-Bellan A, Ferris DK, Vinocour M, Holt JT, Farrar WL. Specific inhibition of c-myc protein biosynthesis using an antisense synthetic deoxy-oligonucleotide in human T lymphocytes. J Immunol. 1988 Apr 1;140(7):2431–2435. [PubMed]
  • Holte H, de Lange Davies C, Beiske K, Stokke T, Marton PF, Smeland EB, Høie J, Kvaløy S. Ki67 and 4F2 antigen expression as well as DNA synthesis predict survival at relapse/tumour progression in low-grade B-cell lymphoma. Int J Cancer. 1989 Dec 15;44(6):975–980. [PubMed]
  • Hwang C, Sinskey AJ, Lodish HF. Oxidized redox state of glutathione in the endoplasmic reticulum. Science. 1992 Sep 11;257(5076):1496–1502. [PubMed]
  • Key G, Meggetto F, Becker MH, al Saati T, Schlüter C, Duchrow M, Delsol G, Gerdes J. Immunobiochemical characterization of the antigen detected by monoclonal antibody IND.64. Evidence that IND.64 reacts with the cell proliferation associated nuclear antigen previously defined by Ki-67. Virchows Arch B Cell Pathol Incl Mol Pathol. 1992;62(4):259–262. [PubMed]
  • Key G, Becker MH, Baron B, Duchrow M, Schlüter C, Flad HD, Gerdes J. New Ki-67-equivalent murine monoclonal antibodies (MIB 1-3) generated against bacterially expressed parts of the Ki-67 cDNA containing three 62 base pair repetitive elements encoding for the Ki-67 epitope. Lab Invest. 1993 Jun;68(6):629–636. [PubMed]
  • Kirschner M. The cell cycle then and now. Trends Biochem Sci. 1992 Aug;17(8):281–285. [PubMed]
  • Krueger NX, Streuli M, Saito H. Structural diversity and evolution of human receptor-like protein tyrosine phosphatases. EMBO J. 1990 Oct;9(10):3241–3252. [PMC free article] [PubMed]
  • Locker AP, Birrell K, Bell JA, Nicholson RI, Elston CW, Blamey RW, Ellis IO. Ki67 immunoreactivity in breast carcinoma: relationships to prognostic variables and short term survival. Eur J Surg Oncol. 1992 Jun;18(3):224–229. [PubMed]
  • McCormick D, Hall PA. The complexities of proliferating cell nuclear antigen. Histopathology. 1992 Dec;21(6):591–594. [PubMed]
  • Nicholson RI, Bouzubar N, Walker KJ, McClelland R, Dixon AR, Robertson JF, Ellis IO, Blamey RW. Hormone sensitivity in breast cancer: influence of heterogeneity of oestrogen receptor expression and cell proliferation. Eur J Cancer. 1991;27(7):908–913. [PubMed]
  • Norbury C, Nurse P. Animal cell cycles and their control. Annu Rev Biochem. 1992;61:441–470. [PubMed]
  • Rechsteiner M. PEST regions, proteolysis and cell cycle progression. Revis Biol Celular. 1989;20:235–253. [PubMed]
  • Rechsteiner M. PEST sequences are signals for rapid intracellular proteolysis. Semin Cell Biol. 1990 Dec;1(6):433–440. [PubMed]
  • Rogers S, Wells R, Rechsteiner M. Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science. 1986 Oct 17;234(4774):364–368. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Saraste M, Sibbald PR, Wittinghofer A. The P-loop--a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci. 1990 Nov;15(11):430–434. [PubMed]
  • Schrape S, Jones DB, Wright DH. A comparison of three methods for the determination of the growth fraction in non-Hodgkin's lymphoma. Br J Cancer. 1987 Mar;55(3):283–286. [PMC free article] [PubMed]
  • Scott RJ, Hall PA, Haldane JS, van Noorden S, Price Y, Lane DP, Wright NA. A comparison of immunohistochemical markers of cell proliferation with experimentally determined growth fraction. J Pathol. 1991 Oct;165(2):173–178. [PubMed]
  • Silver PA. How proteins enter the nucleus. Cell. 1991 Feb 8;64(3):489–497. [PubMed]
  • Toschi L, Bravo R. Changes in cyclin/proliferating cell nuclear antigen distribution during DNA repair synthesis. J Cell Biol. 1988 Nov;107(5):1623–1628. [PMC free article] [PubMed]
  • Triglia T, Peterson MG, Kemp DJ. A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences. Nucleic Acids Res. 1988 Aug 25;16(16):8186–8186. [PMC free article] [PubMed]
  • Tungekar MF, Gatter KC, Dunnill MS, Mason DY. Ki-67 immunostaining and survival in operable lung cancer. Histopathology. 1991 Dec;19(6):545–550. [PubMed]
  • Verheijen R, Kuijpers HJ, Schlingemann RO, Boehmer AL, van Driel R, Brakenhoff GJ, Ramaekers FC. Ki-67 detects a nuclear matrix-associated proliferation-related antigen. I. Intracellular localization during interphase. J Cell Sci. 1989 Jan;92(Pt 1):123–130. [PubMed]
  • Verheijen R, Kuijpers HJ, van Driel R, Beck JL, van Dierendonck JH, Brakenhoff GJ, Ramaekers FC. Ki-67 detects a nuclear matrix-associated proliferation-related antigen. II. Localization in mitotic cells and association with chromosomes. J Cell Sci. 1989 Apr;92(Pt 4):531–540. [PubMed]
  • Wasenius VM, Saraste M, Salvén P, Erämaa M, Holm L, Lehto VP. Primary structure of the brain alpha-spectrin. J Cell Biol. 1989 Jan;108(1):79–93. [PMC free article] [PubMed]
  • Wintzer HO, Zipfel I, Schulte-Mönting J, Hellerich U, von Kleist S. Ki-67 immunostaining in human breast tumors and its relationship to prognosis. Cancer. 1991 Jan 15;67(2):421–428. [PubMed]

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