Skip to main content

Advertisement

SpringerLink
Log in

Development of a novel method for measuring in vivo breast epithelial cell proliferation in humans

  • Report
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Summary

Cell proliferation plays an important role in all stages of carcinogenesis. Currently, no safe, direct, in vivo method of measuring breast epithelial cell (BEC) proliferation rates in humans exists. Static immunohistochemical markers of cell proliferation, such as Ki-67 and PCNA indices, have technical limitations including high inter-lab variability, inaccuracy in the presence of agents that cause G1/S cell cycle block and inadequate sensitivity in post-menopausal women with low BEC proliferation rates. We describe here a safe, direct method of measuring BEC proliferation rates in vivo in women using heavy water (2H2O) labeling coupled with mass spectrometric analysis. Proliferation of normal and tumor BEC was measured from breast tissue biopsies in women undergoing mastectomy (n=11) and normal BEC from healthy volunteers (n=16). Women took heavy water (50–150 ml per day) for 1–4 weeks. Pre-menopausal women had significantly higher proliferation rates than post-menopausal women (0.7 ± 0.1 versus 0.2 ± 0.1 new cells per day, respectively), and tumor BEC had different proliferation rates than normal BEC from the same breast. The method is analytically reproducible and remains sensitive in the range of low proliferation rates. In summary, this novel method of measuring BEC proliferation in vivo holds promise for assessing the effects of anti-proliferative chemopreventive and chemotherapeutic agents.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • DL Page WD Dupont (1992) ArticleTitleBenign breast disease: indicators of increased breast cancer risk Cancer Detect Prev 16 93–97 Occurrence Handle1600526 Occurrence Handle1:STN:280:DyaK383ovVyrtw%3D%3D

    PubMed  CAS  Google Scholar 

  • A Cohen J Barankiewicz HM Lederman EW Gelfand (1983) ArticleTitlePurine and pyrimidine metabolism in human T lymphocytes Regulation of deoxyribonucleotide metabolism. Journal of Biological Chemistry 258 12334–12340 Occurrence Handle1:CAS:528:DyaL2cXjvFGr

    CAS  Google Scholar 

  • PC Clahsen CJ Velde Particlevan de C Duval C Pallud AM Mandard A Delobelle-Deroide L Broek Particlevan den MJ Vijver Particlevan de (1999) ArticleTitleThe utility of mitotic index, oestrogen receptor and Ki-67 measurements in the creation of novel prognostic indices for node-negative breast cancer Eur J Surg Oncol 25 356–363 Occurrence Handle10419704 Occurrence Handle10.1053/ejso.1999.0657 Occurrence Handle1:STN:280:DyaK1MzltValtA%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • P Gaglia A Bernardi T Venesio B Caldarola D Lauro AP Cappa P Calderini DS Liscia (1993) ArticleTitleCell proliferation of breast cancer evaluated by anti-BrdU and anti-Ki-67 antibodies: its prognostic value on short-term recurrences Eur J Cancer 29A 1509–1513 Occurrence Handle8217353 Occurrence Handle10.1016/0959-8049(93)90284-M Occurrence Handle1:STN:280:DyaK2c%2FjtFakuw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • WH Goodson DH Moore BM Ljung K Chew C Florendo B Mayall HS Smith FM Waldman (1998) ArticleTitleThe functional relationship between in vivo bromodeoxyuridine labeling index and Ki-67 proliferation index in human breast cancer Breast Cancer Res Treat 49 155–164 Occurrence Handle9696398 Occurrence Handle10.1023/A:1005926228093 Occurrence Handle1:CAS:528:DyaK1cXkslOiu70%3D

    Article  PubMed  CAS  Google Scholar 

  • WH Goodson SuffixIII DH Moore SuffixII BM Ljung K Chew B Mayall HS Smith FM Waldman (2000) ArticleTitleThe prognostic value of proliferation indices: a study with in vivo bromodeoxyuridine and Ki-67 Breast Cancer Res Treat 59 113–123 Occurrence Handle10817346 Occurrence Handle10.1023/A:1006344010050 Occurrence Handle1:CAS:528:DC%2BD3cXjslWisbc%3D

    Article  PubMed  CAS  Google Scholar 

  • RL Jansen PS Hupperets JW Arends SR Joosten-Achjanie A Volovics HC Schouten HF Hillen (1998) ArticleTitleMIB-1 labelling index is an independent prognostic marker in primary breast cancer Br J Cancer 78 460–465 Occurrence Handle9716027 Occurrence Handle1:STN:280:DyaK1cznvFyrtg%3D%3D

    PubMed  CAS  Google Scholar 

  • P Rudolph P Alm HJ Heidebrecht H Bolte V Ratjen B Baldetorp M Ferno H Olsson R Parwaresch (1999) ArticleTitleImmunologic proliferation marker Ki-S2 as prognostic indicator for lymph node-negative breast cancer J Natl Cancer Inst 91 271–278 Occurrence Handle10037106 Occurrence Handle10.1093/jnci/91.3.271 Occurrence Handle1:STN:280:DyaK1M7lsVOisg%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • AD Thor S Liu DH Moore SuffixII SM Edgerton (1999) ArticleTitleComparison of mitotic index, in vitro bromodeoxyuridine labeling, and MIB-1 assays to quantitate proliferation in breast cancer J Clin Oncol 17 470–477 Occurrence Handle10080587 Occurrence Handle1:STN:280:DyaK1M7nvVKksA%3D%3D

    PubMed  CAS  Google Scholar 

  • M Mengel R Wasielewski Particlevon B Weise T Rudiger H Muller-Hermelink H Kreipe (2002) ArticleTitleInter-laboratory and inter-observer reproducibility of immunohistochemical assessment of the Ki-67 labelling index in a large multi-centre trial J Pathol 198 292–299 Occurrence Handle12375261 Occurrence Handle10.1002/path.1218

    Article  PubMed  Google Scholar 

  • S Cos DE Blask A Lemus-Wilson AB Hill (1991) ArticleTitleEffects of melatonin on the cell cycle kinetics and “estrogen-rescue” of MCF-7 human breast cancer cells in culture J Pineal Res 10 36–42 Occurrence Handle2056430 Occurrence Handle10.1111/j.1600-079X.1991.tb00007.x Occurrence Handle1:CAS:528:DyaK3MXlsFWht7c%3D

    Article  PubMed  CAS  Google Scholar 

  • MP Osborne JF Ruperto JP Crowe PP Rosen NT Telang (1992) ArticleTitleEffect of tamoxifen on preneoplastic cell proliferation in N-nitroso-N-methylurea-induced mammary carcinogenesis Cancer Research 52 1477–1480 Occurrence Handle1540955 Occurrence Handle1:CAS:528:DyaK38XitVOnt78%3D

    PubMed  CAS  Google Scholar 

  • AE Wakeling E Newboult SW Peters (1989) ArticleTitleEffects of antioestrogens on the proliferation of MCF-7 human breast cancer cells J Mol Endocrinol 2 225–234 Occurrence Handle2665780 Occurrence Handle1:CAS:528:DyaL1MXktFOqsLg%3D Occurrence Handle10.1677/jme.0.0020225

    Article  PubMed  CAS  Google Scholar 

  • H Kim TG Peterson S Barnes (1998) ArticleTitleMechanisms of action of the soy isoflavone genistein: emerging role for its effects via transforming growth factor beta signaling pathways Am J Clin Nutri 68 1418S–1425S Occurrence Handle1:CAS:528:DyaK1cXnvV2gurc%3D

    CAS  Google Scholar 

  • E Asher CM Payne C Bernstein (1995) ArticleTitleEvaluation of cell death in EBV-transformed lymphocytes using agarose gel electrophoresis, light microscopy and electron microscopy II. Induction of non-classic apoptosis (“para-apoptosis”) by tritiated thymidine Leukemia and Lymphoma 19 107–119 Occurrence Handle8574155 Occurrence Handle10.3109/10428199509059664 Occurrence Handle1:STN:280:DyaK287hs12hug%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • B Rocha C Penit C Baron F Vasseur N Dautigny AA Freitas (1990) ArticleTitleAccumulation of bromodeoxyuridine-labeled cells in central and peripheral lymphoid organs: minimal estimates of production and turnover rates of mature lymphocytes Euro J Immunol 20 1697–1708 Occurrence Handle10.1002/eji.1830200812 Occurrence Handle1:STN:280:DyaK3M%2Fgs1alug%3D%3D

    Article  CAS  Google Scholar 

  • F Antelo R Neese MK Hellerstein (2000) ArticleTitleA new method for measuring proliferation of adipocytes FASEB 14 A214

    Google Scholar 

  • M Hellerstein MB Hanley D Cesar S Siler C Papageorgopoulos E Wieder D Schmidt R Hoh R Neese D Macallan S Deeks JM McCune (1999) ArticleTitleDirectly measured kinetics of circulating T lymphocytes in normal and HIV-1-infected humans [see comments] Nat Med 5 83–89 Occurrence Handle9883844 Occurrence Handle10.1038/4772 Occurrence Handle1:CAS:528:DyaK1MXmtFOhsg%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • J Kim F Antelo R Neese MK Hellerstein (2000) ArticleTitleA new stable isotope-mass spectrometric (MS) method to measure proliferation rates of colon epithelial cells FASEB 14 A718 Occurrence Handle10.1096/fj.14.11.1508

    Article  Google Scholar 

  • DC Macallan CA Fullerton RA Neese K Haddock SS Park MK Hellerstein (1998) ArticleTitleMeasurement of cell proliferation by labeling of DNA with stable isotope-labeled glucose: studies in vitro, in animals, and in humans Proc Natl Aca Sci US 95 708–713 Occurrence Handle10.1073/pnas.95.2.708 Occurrence Handle1:CAS:528:DyaK1cXmtFaiug%3D%3D

    Article  CAS  Google Scholar 

  • L Misell S Gieng A Gurumurthy M Hellerstein (2002) ArticleTitleGenistein’s effect on mammary epithelial proliferation (MECP) measured in vivo by a stable isotope gas chromatographic/mass spectrometric (GC/MS) method FASEB J 16 A1003

    Google Scholar 

  • L Misell J Thompson F Antelo Y-C Chou S Nandi R Neese M Hellerstein (2000) ArticleTitleA new in vivo stable isotope method for measuring mammary epithelial cell proliferation FASEB J 14 A786

    Google Scholar 

  • LM Misell ES Hwang N Williams MK Hellerstein (2003) ArticleTitleA stable isotope mass spectrometric method for measuring breast epithelial cell proliferation in vivo in women: a promising biomarker for chemoprevention studies Breast Cancer Res Treat 82 s177

    Google Scholar 

  • RA Neese LM Misell S Turner A Chu J Kim D Cesar R Hoh F Antelo A Strawford JM McCune M Christiansen MK Hellerstein (2002) ArticleTitleMeasurement in vivo of proliferation rates of slow turnover cells by 2H2O labeling of the deoxyribose moiety of DNA Proc Natl Acad Sci USA 99 15345–15350 Occurrence Handle12424339 Occurrence Handle10.1073/pnas.232551499 Occurrence Handle1:CAS:528:DC%2BD3sXjvVCm

    Article  PubMed  CAS  Google Scholar 

  • RA Neese SQ Siler D Cesar F Antelo D Lee L Misell K Patel S Tehrani P Shah MK Hellerstein (2001) ArticleTitleAdvances in the stable isotope-mass spectrophotometric measurement of DNA synthesis and cell proliferation Anal Biochem 289 189 Occurrence Handle10.1006/abio.2001.5375 Occurrence Handle1:CAS:528:DC%2BD3MXotFGjtr4%3D

    Article  CAS  Google Scholar 

  • PJ Jones ST Leatherdale (1991) ArticleTitleStable isotopes in clinical research: safety reaffirmed Clin Sci (Lond) 80 277–280 Occurrence Handle1:STN:280:DyaK3M3itlOlug%3D%3D

    CAS  Google Scholar 

  • J Yang R Guzman J Richards V Jentoft MR DeVault SR Wellings S Nandi (1980) ArticleTitlePrimary culture of human mammary epithelial cells embedded in collagen gels J Natl Cancer Inst 65 337–343 Occurrence Handle6995666 Occurrence Handle1:CAS:528:DyaL3cXlt1ygtbs%3D

    PubMed  CAS  Google Scholar 

  • C Clarke J Titley S Davies MJ O’Hare (1994) ArticleTitleAn immunomagnetic separation method using superparamagnetic (MACS) beads for large-scale purification of human mammary luminal and myoepithelial cells Epithelial Cell Biol 3 38–46 Occurrence Handle7514934 Occurrence Handle1:STN:280:DyaK2c3ltFartw%3D%3D

    PubMed  CAS  Google Scholar 

  • JM McCune MB Hanley D Cesar R Halvorsen R Hoh D Schmidt E Wieder S Deeks S Siler R Neese M Hellerstein (2000) ArticleTitleFactors influencing T-cell turnover in HIV-1-seropositive patients [see comments] J Clini Invest 105 R1–8 Occurrence Handle10.1172/JCI8647 Occurrence Handle1:CAS:528:DC%2BD3cXhslygtr0%3D

    Article  CAS  Google Scholar 

  • SF Previs JW Hazey F Diraison M Beylot F David H Brunengraber (1996) ArticleTitleAssay of the deuterium enrichment of water via acetylene J Mass Spectrom 31 639–642 Occurrence Handle8799297 Occurrence Handle10.1002/(SICI)1096-9888(199606)31:6<639::AID-JMS336>3.0.CO;2-S Occurrence Handle1:CAS:528:DyaK28Xjs1Gisrs%3D

    Article  PubMed  CAS  Google Scholar 

  • MK Hellerstein (1999) ArticleTitleMeasurement of T-cell kinetics: recent methodologic advances Immunol Today 20 438–441 Occurrence Handle10500289 Occurrence Handle10.1016/S0167-5699(99)01529-7 Occurrence Handle1:CAS:528:DyaK1MXmsVWhsrw%3D

    Article  PubMed  CAS  Google Scholar 

  • Waterlow JC, Garlick PJ, Millward DJ: Protein Turnover in Mammalian Tissues and in the Whole Body. Amsterdam, 1978

  • K Christov KL Chew BM Ljung FM Waldman LA Duarte WHd Goodson HS Smith BH Mayall (1991) ArticleTitleProliferation of normal breast epithelial cells as shown by in vivo labeling with bromodeoxyuridine American Journal of Pathology 138 1371–1377 Occurrence Handle2053594 Occurrence Handle1:STN:280:DyaK3M3ntlWisA%3D%3D

    PubMed  CAS  Google Scholar 

  • DF Hargreaves F Knox R Swindell CS Potten NJ Bundred (1998) ArticleTitleEpithelial proliferation and hormone receptor status in the normal post-menopausal breast and the effects of hormone replacement therapy Br J Cancer 78 945–949 Occurrence Handle9764588 Occurrence Handle1:CAS:528:DyaK1cXmvVCns7s%3D

    PubMed  CAS  Google Scholar 

  • JS Meyer (1977) ArticleTitleCell proliferation in normal human breast ducts, fibroadenomas, and other ductal hyperplasias measured by nuclear labeling with tritiated thymidine Effects of menstrual phase, age, and oral contraceptive hormones Hum Pathol 8 67–81 Occurrence Handle844855 Occurrence Handle10.1016/S0046-8177(77)80066-X Occurrence Handle1:STN:280:DyaE2s7jvFOjsw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • JS Meyer RE Connor (1982) ArticleTitleCell proliferation in fibrocystic disease and postmenopause breast ducts measured by thymidine labeling Cancer 50 746–751 Occurrence Handle7093909 Occurrence Handle10.1002/1097-0142(19820815)50:4<746::AID-CNCR2820500420>3.0.CO;2-A Occurrence Handle1:STN:280:DyaL383jtlektA%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • NL Petrakis S Barnes EB King J Lowenstein J Wiencke MM Lee R Miike M Kirk L Coward (1996) ArticleTitleStimulatory influence of soy protein isolate on breast secretion in pre- and postmenopausal women Cancer Epidemiol, Biomark Prevent 5 785–794 Occurrence Handle1:CAS:528:DyaK28XmvFKqu74%3D

    CAS  Google Scholar 

  • BS Shoker C Jarvis RB Clarke E Anderson J Hewlett MP Davies DR Sibson JP Sloane (1999) ArticleTitleEstrogen receptor-positive proliferating cells in the normal and precancerous breast Am J Pathol 155 1811–1815 Occurrence Handle10595909 Occurrence Handle1:CAS:528:DC%2BD3cXit1Kiuw%3D%3D

    PubMed  CAS  Google Scholar 

  • KJ Walker JM Price-Thomas W Candlish RI Nicholson (1991) ArticleTitleInfluence of the antioestrogen tamoxifen on normal breast tissue Br J Cancer 64 764–768 Occurrence Handle1911227 Occurrence Handle1:STN:280:DyaK38%2FgtFeitA%3D%3D

    PubMed  CAS  Google Scholar 

  • J Russo G Calaf L Roi IH Russo (1987) ArticleTitleInfluence of age and gland topography on cell kinetics of normal human breast tissue J Natl Cancer Inst 78 413–418 Occurrence Handle3469454 Occurrence Handle1:STN:280:DyaL2s7kt1Cmtg%3D%3D

    PubMed  CAS  Google Scholar 

  • MC Pike DV Spicer L Dahmoush MF Press (1993) ArticleTitleEstrogens, progestogens, normal breast cell proliferation, and breast cancer risk Epidemiol Rev 15 17–35 Occurrence Handle8405201 Occurrence Handle1:STN:280:DyaK2c%2FgvFCjsQ%3D%3D

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. KineMed, Inc., 5980 Horton St., Suite 470, Emeryville, CA, 94608

    Lisa M. Misell

  2. Carol Franc Buck Breast Care Clinic, Comprehensive Cancer Center, University of California, San Francisco

    E. Shelley Hwang, Alfred Au & Laura Esserman

  3. Division of Endocrinology and Metabolism, Department of Medicine, University of California, at San Francisco, San Francisco

    Marc K. Hellerstein

  4. Department of Nutritional Sciences, University of California at Berkeley, CA, USA

    Marc K. Hellerstein

Authors
  1. Lisa M. Misell
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Shelley Hwang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alfred Au
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Laura Esserman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marc K. Hellerstein
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lisa M. Misell.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Misell, L.M., Hwang, E.S., Au, A. et al. Development of a novel method for measuring in vivo breast epithelial cell proliferation in humans. Breast Cancer Res Treat 89, 257–264 (2005). https://doi.org/10.1007/s10549-004-2228-5

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-004-2228-5

Keywords

Search

Navigation