Advertisement

Abstract

Mapping of homozygous deletions on human chromosome 10q23 has led to the isolation of a candidate tumor suppressor gene, PTEN, that appears to be mutated at considerable frequency in human cancers. In preliminary screens, mutations of PTEN were detected in 31% (13/42) of glioblastoma cell lines and xenografts, 100% (4/4) of prostate cancer cell lines, 6% (4/65) of breast cancer cell lines and xenografts, and 17% (3/18) of primary glioblastomas. The predicted PTEN product has a protein tyrosine phosphatase domain and extensive homology to tensin, a protein that interacts with actin filaments at focal adhesions. These homologies suggest that PTEN may suppress tumor cell growth by antagonizing protein tyrosine kinases and may regulate tumor cell invasion and metastasis through interactions at focal adhesions.
Get full access to this article

View all available purchase options and get full access to this article.

Already a Subscriber?

REFERENCES AND NOTES

1.
Bigner S. H., et al., Cancer Res. 48, 405 (1988)C. D. James et al., ibid., p. 5546; B. K. A. Rasheed, et al., Oncogene 10, 2243 (1995).
2.
Gray I. C., et al., Cancer Res. 55, 4800 (1995)M. Ittmann, ibid. 56, 2143 (1996); T. Trybus, A. Burgess, K. Wojno, T. Glover, J. Macoska, ibid., p. 2263.
3.
Hsu S., et al., ibid. 56, 5684 (1996).
4.
Lisitsyn N. A., Wigler M., Science 259, 946 (1993)M. Schutte, et al., Proc. Natl. Acad. Sci. U.S.A. 92, 5950 (1995); RDA was performed as described in N. A. Lisitsyn et al. (ibid., p. 151). Diploid and aneuploid nuclei from primary breast cancer cells were separated with a fluorescence-activated cell sorter. DNA (100 ng) from each fraction was digested with Bgl II and used to prepare amplicons for 12 separate RDA reactions. Probe CY17 was isolated from one of these reactions. CY17 was 236 bp long and was present in the diploid but not in the aneuploid amplicon from which it was derived. Hybridization of CY17 to normal genomic DNA samples digested with Bgl II revealed no evidence of restriction length polymorphism.
5.
Cox D. R., Burmeister M., Price E. R., Kim S., Myers R. M., Science 250, 245 (1990)We generated primers to amplify CY17 and screened the GeneBridge4 radiation hybrid panel. The primers were 5′-ATCTAGTGAGTTGGGGGACAGAGG-3′ and 5′-CTGGGTTAGGGATTCTGCTCAG-3′. Amplification conditions were 95°C for 30 s, 56°C for 1 min, and 70°C for 1 min for 35 cycles.
6.
Hudson T. J., et al., Science 270, 1945 (1995).
7.
The CEPH (Centre d'Etude du Polymorphisme Humain) B library (Research Genetics, Huntsville, AL) was screened by PCR using a series of tiered pools to identify unique clones.
8.
The forward PCR primer was labeled with [32P]ATP and used to amplify 40 ng of genomic DNA. The samples were then subjected to electrophoresis and autoradiography. The samples included 25 human breast tumor cell lines available from American Type Culture Collection (ATCC) as well as 40 human primary breast tumors xenografted into nude mice. The cell lines were HS578T, SK-BR-3, UACC812, UACC893, MDA-MB-453, MDA-MB-175-VII, MDA-MB468, MDA-MB-361, MDA-MB-231, MDA-MB-436, MDA-MB-415, MDA-MB-330, MDA-MB-157, MDA-MB-134-VI, MDA-MB-435S, ZR 75-30, ZR 75-1, BT-549, BT-483, T-47D, BT-474, DU4475, CAMA1, MCF7, and BT-20.
9.
Kim U., et al., Genomics 34, 213 (1996)Clones were isolated from a BAC library (Research Genetics) using AFMA086WG9 as an STS probe.
10.
BAC DNA was prepared using the Nucleobond kit (Nest Group, Southboro, MA). BACs were digested with Not I and subjected to electrophoresis on a field inversion apparatus. BACs A, B, C, and D were 240, 200, 175, and 120 kb, respectively (see Fig. 1B). A Not I site was present 20 kb from one end of BAC D. Twelve new STS sites were generated by sequencing both ends of BACs B, C, and D, and shotgun cloning Eco RI fragments. Plasmid DNA was prepared from the cloned Eco RI fragments. DNA was cycle sequenced with appropriate primers using a [33P]ddNTP cycle sequencing kit (Amersham). STS primers were designed and the relative location of the STSs determined by testing for their presence in the BAC contig. Primer sequences are available upon request.
11.
The glioblastoma lines included U105, U118MG, A172, DBTRG-05MG, U373MG, T-98G, U-87MG, and U138MG and 34 glioblastoma xenografts. The prostate cancer cell lines tested were DU145, LNCaP, NCI H660, and PC-3, and microsatellite analysis revealed that each was unique. With the exception of U105, all lines were obtained from ATCC.
12.
DNA (10 μg) was digested with Eco RI, resolved on a 1% agarose gel, and transferred to nylon. The JL25 3-kb probe and the 2-kb control probe were randomly labeled and hybridized to the blot consecutively.
13.
Buckler A. J., et al., Proc. Natl. Acad. Sci. U.S.A. 88, 4005 (1991)
Lennon G., Auffray C., Polymeropoulos M., Soares M. B., Genomics 33, 151 (1996).
14.
The map panel #2 monochromosome panel was purchased from the National Institute of General Medical Science (NIGMS) Human Mutant Genetic Cell Repository.
15.
Tonks N. K., Neel B. G., Cell 87, 365 (1996).
16.
Fauman E. B., Saper M. A., Trends Biochem. Sci. 21, 413 (1996);
Sheng Z. and, Charboneau H., J. Biol. Chem. 268, 4728 (1993).
17.
Diamond R. H., Cressman D. E., Laz T. M., Abrams C. S., Taub R., Mol. Cell. Biol. 14, 3752 (1994);
Hogan E. and , Koshland D., Proc. Natl. Acad. Sci. U.S.A. 89, 3098 (1992).
18.
Haynie D. T., Ponting C. P., Protein Sci. 5, 2643 (1996).
19.
Denu J. M., Stuckey J. A., Saper M. A., Dixon J. E., Cell 87, 361 (1996).
20.
Powell S. M., et al., N. Engl. J. Med. 329, 1982 (1993);
Roest P. A. M., Roberts R. G., Sugino S., van Ommen J. B., den Dunnen J. T., Hum. Mol. Genet. 2, 1719 (1993).
21.
STSs Not-5′, PTPD, and ET-1 were amplified from primary glioblastoma DNA and blood DNA and the exonic regions were sequenced.
22.
Hunter T., Cell 50, 823 (1987).
23.
Wilkins J. A., Risinger M. A., Lin S., J. Cell Biol. 103, 1483 (1986)J. Z. Chuang, D. C. Lin, S. Lin, ibid. 128, 1095 (1995); S. Miyamoto, et al., ibid. 131, 791 (1995); S. Miyamoto, S. K. Akiyama, K. M. Yamada, Science 267, 883 (1995).
24.
Zhu X., Ohtsubo M., Bohmer R. M., Roberts J. M., Assoian R. K., J. Cell Biol. 133, 391 (1996);
Wary K. K., Mainiero F., Isakoff S. J., Marcantonio E. E., Giancotti F. E., Cell 87, 733 (1996).
25.
Akiyama S. K., Olden K., Yamada K. M., Cancer Metastasis Rev. 14, 173 (1995).
26.
We thank B. Vogelstein, N. Tonks, and E. Marcantonio for their comments and S. Kalachikov and R. Hauptschein for helpful suggestions. R.P. is a James S. McDonnell Scholar. M.H.W. is an American Cancer Society Research Professor and is supported by the Department of the Army (DAMD 17-94-I4247), NCI (5R35 CA39829), Amplicon Corporation, and the “1 in 9” breast cancer organization. This work is dedicated to the memory of Richard K. Parsons and Richard P. Sanchez.

Information & Authors

Information

Published In

Science
Volume 275 | Issue 5308
28 March 1997

Submission history

Received: 3 February 1997
Accepted: 27 February 1997
Published in print: 28 March 1997

Permissions

Request permissions for this article.

Authors

Affiliations

Jing Li*
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.
Clifford Yen*
C. Yen, L. Rodgers, R. McCombie, M. Wigler, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Danny Liaw*
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.
Katrina Podsypanina*
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.
Shikha Bose
S. Bose, B. Tycko, H. Hibshoosh, Department of Pathology, College of Physicians & Surgeons, Columbia University, New York, NY 10032, USA.
Steven I. Wang
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.
Janusz Puc
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.
Christa Miliaresis
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.
Linda Rodgers
C. Yen, L. Rodgers, R. McCombie, M. Wigler, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Richard McCombie
C. Yen, L. Rodgers, R. McCombie, M. Wigler, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Sandra H. Bigner
S. H. Bigner, Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA.
Beppino C. Giovanella
B. Giovanella, Stehlin Foundation for Cancer Research, St. Joseph Hospital, Houston, TX 77003, USA.
Michael Ittmann
M. Ittmann, New York VA Medical Center and Department of Pathology, New York University, 423 East 23 Street, New York, NY 10010, USA.
Ben Tycko
S. Bose, B. Tycko, H. Hibshoosh, Department of Pathology, College of Physicians & Surgeons, Columbia University, New York, NY 10032, USA.
Hanina Hibshoosh
S. Bose, B. Tycko, H. Hibshoosh, Department of Pathology, College of Physicians & Surgeons, Columbia University, New York, NY 10032, USA.
Michael H. Wigler
C. Yen, L. Rodgers, R. McCombie, M. Wigler, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Ramon Parsons
J. Li, D. Liaw, K. Podsypanina, S. I. Wang, J. Puc, C. Miliaresis, R. Parsons, Department of Pathology and Department of Medicine, College of Physicians & Surgeons, Columbia University, 630 West 168 Street, New York, NY 10032, USA.

Notes

* These authors contributed equally to this work.
† To whom correspondence should be addressed.

Metrics & Citations

Metrics

Article Usage
Altmetrics

Citations

Export citation

Select the format you want to export the citation of this publication.

Cited by
  1. Mechanisms of stearoyl CoA desaturase inhibitor sensitivity and acquired resistance in cancer, Science Advances, 7, 7, (2021)./doi/10.1126/sciadv.abd7459
    Abstract
  2. PTEN status determines chemosensitivity to proteasome inhibition in cholangiocarcinoma, Science Translational Medicine, 12, 562, (2021)./doi/10.1126/scitranslmed.aay0152
    Abstract
  3. Restoring tumor suppression, Science, 364, 6441, (633-634), (2021)./doi/10.1126/science.aax5526
    Abstract
  4. Reactivation of PTEN tumor suppressor for cancer treatment through inhibition of a MYC-WWP1 inhibitory pathway, Science, 364, 6441, (2021)./doi/10.1126/science.aau0159
    Abstract
  5. Skp2-dependent reactivation of AKT drives resistance to PI3K inhibitors, Science Signaling, 11, 521, (2021)./doi/10.1126/scisignal.aao3810
    Abstract
  6. Rb1 and Trp53 cooperate to suppress prostate cancer lineage plasticity, metastasis, and antiandrogen resistance, Science, 355, 6320, (78-83), (2021)./doi/10.1126/science.aah4199
    Abstract
  7. Synaptojanin 2 is a druggable mediator of metastasis and the gene is overexpressed and amplified in breast cancer, Science Signaling, 8, 360, (ra7-ra7), (2021)./doi/10.1126/scisignal.2005537
    Abstract
  8. A Unified Nomenclature and Amino Acid Numbering for Human PTEN, Science Signaling, 7, 332, (pe15-pe15), (2021)./doi/10.1126/scisignal.2005560
    Abstract
  9. A Secreted PTEN Phosphatase That Enters Cells to Alter Signaling and Survival, Science, 341, 6144, (399-402), (2021)./doi/10.1126/science.1234907
    Abstract
  10. Network Analysis of the Focal Adhesion to Invadopodia Transition Identifies a PI3K-PKCα Invasive Signaling Axis, Science Signaling, 5, 241, (ra66-ra66), (2021)./doi/10.1126/scisignal.2002964
    Abstract
Loading...

View Options

Get Access

Log in to view the full text

AAAS Log in

AAAS login provides access to Science for AAAS members, and access to other journals in the Science family to users who have purchased individual subscriptions.

Log in via OpenAthens.
Log in via Shibboleth.
More options

Purchase digital access to this article

Download and print this article for your personal scholarly, research, and educational use.

Purchase this issue in print

Buy a single issue of Science for just $15 USD.

View options

PDF format

Download this article as a PDF file

Download PDF

Media

Figures

Multimedia

Tables

Share

Share

Share article link

Share on social media