DOI: 10.1073/pnas.0603672103. Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness

Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness

10.1073/pnas.0603672103

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The recent identification of “side population” (SP) cells in a number of unrelated human cancers and their normal tissue sources has renewed interest in the hypothesis that cancers may arise from somatic stem/progenitor cells. The high incidence of recurrence attributable to multidrug resistance and the multiple histologic phenotypes indicative of multipotency suggests a stem cell-like etiology of ovarian cancer. Here we identify and characterize SP cells from two distinct genetically engineered mouse ovarian cancer cell lines. Differential efflux of the DNA-binding dye Hoechst 33342 from these cell lines defined a human breast cancer-resistance protein 1-expressing, verapamil-sensitive SP of candidate cancer stem cells.In vivo, mouse SP cells formed measurable tumors sooner than non-SP (NSP) cells when equal numbers were injected into the dorsal fat pad of nude mice. The presence of Mullerian Inhibiting Substance (MIS) signaling pathway transduction molecules in both SP and NSP mouse cells led us to investigate the efficacy of MIS against these populations in comparison with traditional chemotherapies. MIS inhibited the proliferation of both SP and NSP cells, whereas the lipophilic chemotherapeutic agent doxorubicin more significantly inhibited the NSP cells. Finally, we identified breast cancer-resistance protein 1-expressing verapamil-sensitive SPs in three of four human ovarian cancer cell lines and four of six patient primary ascites cells. In the future, individualized therapy must incorporate analysis of the stem cell-like subpopulation of ovarian cancer cells when designing therapeutic strategies for ovarian cancer patients.

Bibliography

Szotek, P. P., Pieretti-Vanmarcke, R., Masiakos, P. T., Dinulescu, D. M., Connolly, D., Foster, R., Dombkowski, D., Preffer, F., MacLaughlin, D. T., & Donahoe, P. K. (2006). Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness. Proceedings of the National Academy of Sciences, 103(30), 11154â11159.

Authors 10

Paul P. Szotek (first)
Rafael Pieretti-Vanmarcke (additional)
Peter T. Masiakos (additional)
Daniela M. Dinulescu (additional)
Denise Connolly (additional)
Rosemary Foster (additional)
David Dombkowski (additional)
Frederic Preffer (additional)
David T. MacLaughlin (additional)
Patricia K. Donahoe (additional)

References 42 Referenced 580

10.1073/pnas.0530291100
10.1634/stemcells.2005-0282
10.1073/pnas.0307618100
10.1182/blood.V98.4.1166
G. M. Seigel, L. M. Campbell, M. Narayan, F. Gonzalez-Fernandez Mol. Vis. 11, 729–737 (2005). / Mol. Vis. by Seigel G. M. (2005)
G. Poste, R. Greig Invasion Metastasis 2, 137–176 (1982). / Invasion Metastasis by Poste G. (1982)
10.1073/pnas.79.21.6574
10.1038/35102167
10.1016/S1470-2045(02)00820-3
10.1146/annurev.cellbio.21.012704.131525
10.1186/1477-7827-2-20
10.1016/S0046-8177(70)80005-3
10.1038/modpathol.3800305
10.3322/canjclin.55.1.10
10.1056/NEJM199311183292108
10.3322/canjclin.47.1.5
10.1038/nrc706
10.1084/jem.183.4.1797
D. C. Connolly, R. Bao, A. Y. Nikitin, K. C. Stephens, T. W. Poole, X. Hua, S. S. Harris, B. C. Vanderhyden, T. C. Hamilton Cancer Res. 63, 1389–1397 (2003). / Cancer Res. by Connolly D. C. (2003)
10.1038/nm1173
10.1073/pnas.0400067101
10.1038/sj.onc.1206938
10.1146/annurev.bi.62.070193.002125
10.1016/j.critrevonc.2004.04.007
10.1158/1078-0432.CCR-05-2108
10.1242/dev.02383
10.1093/carcin/bgh293
R. P. Gale, A. Butturini Leukemia 6, 80–85 (1992). / Leukemia by Gale R. P. (1992)
10.1111/j.1749-6632.1956.tb50894.x
Z. Duan, K. A. Brakora, M. V. Seiden Mol. Cancer Ther. 3, 833–838 (2004). (10.1158/1535-7163.833.3.7) / Mol. Cancer Ther. by Duan Z. (2004)
Z. Duan, D. E. Lamendola, R. Z. Yusuf, R. T. Penson, F. I. Preffer, M. V. Seiden Anticancer Res. 22, 1933–1941 (2002). / Anticancer Res. by Duan Z. (2002)
10.1016/j.ygyno.2003.11.053
10.1006/gyno.2002.6762
10.1016/0090-8258(85)90019-8
10.1074/jbc.M005701200
10.1634/stemcells.20-5-417
10.1002/ijc.2910450306
10.1016/0022-1759(86)90368-6
H. K. Lorenzo, J. Teixeira, N. Pahlavan, V. M. Laurich, P. K. Donahoe, D. T. MacLaughlin J. Chromatogr. B Biomed. Sci. Appl. 766, 89–98 (2002). / J. Chromatogr. B Biomed. Sci. Appl. by Lorenzo H. K. (2002)
10.1210/jcem-70-1-16
10.1095/biolreprod15.3.329
10.1016/0022-4804(77)90202-5

Dates

Type	When
Created	19 years, 1 month ago (July 18, 2006, 8:35 p.m.)
Deposited	2 years, 3 months ago (May 8, 2023, 12:11 a.m.)
Indexed	10 hours, 52 minutes ago (Sept. 3, 2025, 6:48 a.m.)
Issued	19 years, 1 month ago (July 25, 2006)
Published	19 years, 1 month ago (July 25, 2006)
Published Online	19 years, 1 month ago (July 25, 2006)
Published Print	19 years, 1 month ago (July 25, 2006)

Funders 0

None

BibTeX

@article{Szotek_2006, title={Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness}, volume={103}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0603672103}, DOI={10.1073/pnas.0603672103}, number={30}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Szotek, Paul P. and Pieretti-Vanmarcke, Rafael and Masiakos, Peter T. and Dinulescu, Daniela M. and Connolly, Denise and Foster, Rosemary and Dombkowski, David and Preffer, Frederic and MacLaughlin, David T. and Donahoe, Patricia K.}, year={2006}, month=jul, pages={11154–11159} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        9,
        3
      ]
    ],
    "date-time": "2025-09-03T10:48:40Z",
    "timestamp": 1756896520758
  },
  "reference-count": 42,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "30",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2006,
        7,
        25
      ]
    ]
  },
  "abstract": "<jats:p>The recent identification of \u201cside population\u201d (SP) cells in a number of unrelated human cancers and their normal tissue sources has renewed interest in the hypothesis that cancers may arise from somatic stem/progenitor cells. The high incidence of recurrence attributable to multidrug resistance and the multiple histologic phenotypes indicative of multipotency suggests a stem cell-like etiology of ovarian cancer. Here we identify and characterize SP cells from two distinct genetically engineered mouse ovarian cancer cell lines. Differential efflux of the DNA-binding dye Hoechst 33342 from these cell lines defined a human breast cancer-resistance protein 1-expressing, verapamil-sensitive SP of candidate cancer stem cells.<jats:italic>In vivo</jats:italic>, mouse SP cells formed measurable tumors sooner than non-SP (NSP) cells when equal numbers were injected into the dorsal fat pad of nude mice. The presence of Mullerian Inhibiting Substance (MIS) signaling pathway transduction molecules in both SP and NSP mouse cells led us to investigate the efficacy of MIS against these populations in comparison with traditional chemotherapies. MIS inhibited the proliferation of both SP and NSP cells, whereas the lipophilic chemotherapeutic agent doxorubicin more significantly inhibited the NSP cells. Finally, we identified breast cancer-resistance protein 1-expressing verapamil-sensitive SPs in three of four human ovarian cancer cell lines and four of six patient primary ascites cells. In the future, individualized therapy must incorporate analysis of the stem cell-like subpopulation of ovarian cancer cells when designing therapeutic strategies for ovarian cancer patients.</jats:p>",
  "DOI": "10.1073/pnas.0603672103",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        7,
        19
      ]
    ],
    "date-time": "2006-07-19T00:35:07Z",
    "timestamp": 1153269307000
  },
  "page": "11154-11159",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 580,
  "title": "Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness",
  "prefix": "10.1073",
  "volume": "103",
  "author": [
    {
      "given": "Paul P.",
      "family": "Szotek",
      "sequence": "first",
      "affiliation": [
        {
          "name": "*Pediatric Surgical Research Laboratories, Department of Surgery, and"
        }
      ]
    },
    {
      "given": "Rafael",
      "family": "Pieretti-Vanmarcke",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "*Pediatric Surgical Research Laboratories, Department of Surgery, and"
        }
      ]
    },
    {
      "given": "Peter T.",
      "family": "Masiakos",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "*Pediatric Surgical Research Laboratories, Department of Surgery, and"
        }
      ]
    },
    {
      "given": "Daniela M.",
      "family": "Dinulescu",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Pathology, Eugene Braunwald Research Center, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Room 401a, Boston, MA 02115; and"
        }
      ]
    },
    {
      "given": "Denise",
      "family": "Connolly",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, PA 19111;"
        }
      ]
    },
    {
      "given": "Rosemary",
      "family": "Foster",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Medicine, Division of Hematology/Oncology, Massachusetts General Hospital, Harvard Medical School, 70 Blossom Street, Boston, MA 02114"
        }
      ]
    },
    {
      "given": "David",
      "family": "Dombkowski",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Flow Cytometry Laboratory, Department of Pathology and Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Medical School, 185 Cambridge Street, Boston, MA 02114;"
        }
      ]
    },
    {
      "given": "Frederic",
      "family": "Preffer",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Flow Cytometry Laboratory, Department of Pathology and Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Medical School, 185 Cambridge Street, Boston, MA 02114;"
        }
      ]
    },
    {
      "given": "David T.",
      "family": "MacLaughlin",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "*Pediatric Surgical Research Laboratories, Department of Surgery, and"
        }
      ]
    },
    {
      "given": "Patricia K.",
      "family": "Donahoe",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "*Pediatric Surgical Research Laboratories, Department of Surgery, and"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        2006,
        7,
        25
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_4_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0530291100"
    },
    {
      "key": "e_1_3_4_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1634/stemcells.2005-0282"
    },
    {
      "key": "e_1_3_4_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0307618100"
    },
    {
      "key": "e_1_3_4_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1182/blood.V98.4.1166"
    },
    {
      "key": "e_1_3_4_5_2",
      "first-page": "729",
      "volume": "11",
      "author": "Seigel G. M.",
      "year": "2005",
      "unstructured": "G. M. Seigel, L. M. Campbell, M. Narayan, F. Gonzalez-Fernandez Mol. Vis. 11, 729\u2013737 (2005).",
      "journal-title": "Mol. Vis."
    },
    {
      "key": "e_1_3_4_6_2",
      "first-page": "137",
      "volume": "2",
      "author": "Poste G.",
      "year": "1982",
      "unstructured": "G. Poste, R. Greig Invasion Metastasis 2, 137\u2013176 (1982).",
      "journal-title": "Invasion Metastasis"
    },
    {
      "key": "e_1_3_4_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.79.21.6574"
    },
    {
      "key": "e_1_3_4_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/35102167"
    },
    {
      "key": "e_1_3_4_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1470-2045(02)00820-3"
    },
    {
      "key": "e_1_3_4_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.cellbio.21.012704.131525"
    },
    {
      "key": "e_1_3_4_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1186/1477-7827-2-20"
    },
    {
      "key": "e_1_3_4_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0046-8177(70)80005-3"
    },
    {
      "key": "e_1_3_4_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/modpathol.3800305"
    },
    {
      "key": "e_1_3_4_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.3322/canjclin.55.1.10"
    },
    {
      "key": "e_1_3_4_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1056/NEJM199311183292108"
    },
    {
      "key": "e_1_3_4_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.3322/canjclin.47.1.5"
    },
    {
      "key": "e_1_3_4_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nrc706"
    },
    {
      "key": "e_1_3_4_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1084/jem.183.4.1797"
    },
    {
      "key": "e_1_3_4_19_2",
      "first-page": "1389",
      "volume": "63",
      "author": "Connolly D. C.",
      "year": "2003",
      "unstructured": "D. C. Connolly, R. Bao, A. Y. Nikitin, K. C. Stephens, T. W. Poole, X. Hua, S. S. Harris, B. C. Vanderhyden, T. C. Hamilton Cancer Res. 63, 1389\u20131397 (2003).",
      "journal-title": "Cancer Res."
    },
    {
      "key": "e_1_3_4_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nm1173"
    },
    {
      "key": "e_1_3_4_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0400067101"
    },
    {
      "key": "e_1_3_4_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/sj.onc.1206938"
    },
    {
      "key": "e_1_3_4_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.bi.62.070193.002125"
    },
    {
      "key": "e_1_3_4_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.critrevonc.2004.04.007"
    },
    {
      "key": "e_1_3_4_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1158/1078-0432.CCR-05-2108"
    },
    {
      "key": "e_1_3_4_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1242/dev.02383"
    },
    {
      "key": "e_1_3_4_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/carcin/bgh293"
    },
    {
      "key": "e_1_3_4_28_2",
      "first-page": "80",
      "volume": "6",
      "author": "Gale R. P.",
      "year": "1992",
      "unstructured": "R. P. Gale, A. Butturini Leukemia 6, 80\u201385 (1992).",
      "journal-title": "Leukemia"
    },
    {
      "key": "e_1_3_4_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1749-6632.1956.tb50894.x"
    },
    {
      "key": "e_1_3_4_30_2",
      "doi-asserted-by": "crossref",
      "first-page": "833",
      "DOI": "10.1158/1535-7163.833.3.7",
      "volume": "3",
      "author": "Duan Z.",
      "year": "2004",
      "unstructured": "Z. Duan, K. A. Brakora, M. V. Seiden Mol. Cancer Ther. 3, 833\u2013838 (2004).",
      "journal-title": "Mol. Cancer Ther."
    },
    {
      "key": "e_1_3_4_31_2",
      "first-page": "1933",
      "volume": "22",
      "author": "Duan Z.",
      "year": "2002",
      "unstructured": "Z. Duan, D. E. Lamendola, R. Z. Yusuf, R. T. Penson, F. I. Preffer, M. V. Seiden Anticancer Res. 22, 1933\u20131941 (2002).",
      "journal-title": "Anticancer Res."
    },
    {
      "key": "e_1_3_4_32_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.ygyno.2003.11.053"
    },
    {
      "key": "e_1_3_4_33_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/gyno.2002.6762"
    },
    {
      "key": "e_1_3_4_34_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0090-8258(85)90019-8"
    },
    {
      "key": "e_1_3_4_35_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M005701200"
    },
    {
      "key": "e_1_3_4_36_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1634/stemcells.20-5-417"
    },
    {
      "key": "e_1_3_4_37_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/ijc.2910450306"
    },
    {
      "key": "e_1_3_4_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-1759(86)90368-6"
    },
    {
      "key": "e_1_3_4_39_2",
      "first-page": "89",
      "volume": "766",
      "author": "Lorenzo H. K.",
      "year": "2002",
      "unstructured": "H. K. Lorenzo, J. Teixeira, N. Pahlavan, V. M. Laurich, P. K. Donahoe, D. T. MacLaughlin J. Chromatogr. B Biomed. Sci. Appl. 766, 89\u201398 (2002).",
      "journal-title": "J. Chromatogr. B Biomed. Sci. Appl."
    },
    {
      "key": "e_1_3_4_40_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1210/jcem-70-1-16"
    },
    {
      "key": "e_1_3_4_41_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1095/biolreprod15.3.329"
    },
    {
      "key": "e_1_3_4_42_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-4804(77)90202-5"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.0603672103",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        5,
        8
      ]
    ],
    "date-time": "2023-05-08T04:11:29Z",
    "timestamp": 1683519089000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.0603672103"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2006,
        7,
        25
      ]
    ]
  },
  "references-count": 42,
  "journal-issue": {
    "issue": "30",
    "published-print": {
      "date-parts": [
        [
          2006,
          7,
          25
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.0603672103"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.0603672103",
  "relation": {
    "has-review": [
      {
        "id-type": "doi",
        "id": "10.3410/f.1081958.534905",
        "asserted-by": "object"
      }
    ]
  },
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        2006,
        7,
        25
      ]
    ]
  },
  "assertion": [
    {
      "value": "2006-07-25",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}