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. 2015 May 28:5:10357.
doi: 10.1038/srep10357.

Identification of a distinct population of CD133(+)CXCR4(+) cancer stem cells in ovarian cancer

Michele Cioffi  1 , Crescenzo D'Alterio  1 , Rosalba Camerlingo  2 , Virginia Tirino  2 , Claudia Consales  1 , Anna Riccio  1 , Caterina Ieranò  1 , Sabrina Chiara Cecere  3 , Nunzia Simona Losito  4 , Stefano Greggi  3 , Sandro Pignata  3 , Giuseppe Pirozzi  2 , Stefania Scala  1
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Identification of a distinct population of CD133(+)CXCR4(+) cancer stem cells in ovarian cancer

Michele Cioffi et al. Sci Rep. .

Abstract

CD133 and CXCR4 were evaluated in the NCI-60 cell lines to identify cancer stem cell rich populations. Screening revealed that, ovarian OVCAR-3, -4 and -5 and colon cancer HT-29, HCT-116 and SW620 over expressed both proteins. We aimed to isolate cells with stem cell features sorting the cells expressing CXCR4(+)CD133(+) within ovarian cancer cell lines. The sorted population CD133(+)CXCR4(+) demonstrated the highest efficiency in sphere formation in OVCAR-3, OVCAR-4 and OVCAR-5 cells. Moreover OCT4, SOX2, KLF4 and NANOG were highly expressed in CD133(+)CXCR4(+) sorted OVCAR-5 cells. Most strikingly CXCR4(+)CD133(+) sorted OVCAR-5 and -4 cells formed the highest number of tumors when inoculated in nude mice compared to CD133(-)CXCR4(-), CD133(+)CXCR4(-), CD133(-)CXCR4(+) cells. CXCR4(+)CD133(+) OVCAR-5 cells were resistant to cisplatin, overexpressed the ABCG2 surface drug transporter and migrated toward the CXCR4 ligand, CXCL12. Moreover, when human ovarian cancer cells were isolated from 37 primary ovarian cancer, an extremely variable level of CXCR4 and CD133 expression was detected. Thus, in human ovarian cancer cells CXCR4 and CD133 expression identified a discrete population with stem cell properties that regulated tumor development and chemo resistance. This cell population represents a potential therapeutic target.

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Figures

Figure 1
Figure 1. CXCR4 and CD133 protein expression in NCI 60 cell lines.
(A) CXCR4 and CD133 protein level was evaluated through Immunoblotting. All gels had been run under the same experimental conditions. (B) Bar plots summarizing Flow Cytometry analysis for CXCR4 and CD133 protein level in the 60 cell lines from the Drug Screen Program.
Figure 2
Figure 2. CXCR4+CD133+ ovarian cancer cells show stem cell properties.
(A) For in vitro and in vivo experiments, cells were double-stained for CD133 and CXCR4. Four distinct phenotypic subpopulations, specifically CD133−CXCR4−, CD133−CXCR4+, CD133+CXCR4− and CD133+CXCR4+, were isolated. (B) Sphere formation capacity of sorted population in OVCAR-3, OVCAR-4 and OVCAR-5 cells. Tumor spheroids under non-differentiating and non-adherent conditions are known to contain a greater number of CSCs, images of OVCAR-5 sphere as representative of anchorage-independent growth, and tumor spheroid formation was reported. (Left side, B); Bar graph depicting number of spheres observed in sorted cells culture (Right side, B); QPCR analysis of pluripotency-associated genes (OCT4, SOX2, KLF4, NANOG) in OVCAR-5 sorted populations (C). The data represent the mean ± SD. Asterisk (*) represents p values < 0.05; double asterisk (**) represents p values < 0.001.
Figure 3
Figure 3. CXCR4+CD133+ ovarian cancer cells possess resistance to chemotherapy, migration and colony forming capabilities.
OVCAR-5 sorted cells (A) Cytotoxicity assay in the presence of Cisplatin (0, 5, 10M). (B) QPCR analysis of ABCG2. (C) Migration assay toward CXCL12 and (D) Clonogenic assay. A representative photograph is provided in the lower panel. (*p < 0.05, **p values < 0.001, compared with the CD133-CXCR4- group).
Figure 4
Figure 4. CXCR4 and CD133 are highly expressed in ovarian cancer patients.
(A) Representative panel of immunohistochemistry for CD133 and CXCR4 expression (Magnification 200X; insert 400X). Non-homogeneous and focal cytoplasmic and membranous CD133 expression rated as negative (I), focally low (1–10% cancer cells) (II), focally high CD133 expression (>10% stained cancer cells) (III). Extensive homogeneous cytoplasmic and membranous, CXCR4 staining was reported rated as negative (IV) moderate (<50% of cancer cells) (V) and high expression (>50% of cancer cells) (VI). (B) QPCR analysis of CXCR4 and CD133 in freshly resected ovarian tumors and corresponding normal ovarian tissue.
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References

    1. Visvader J.E. & Lindeman G.J. Cancer stem cells in solid tumors: accumulating evidence and unresolved questions. Nat. Rev. Cancer 8, 755–768 (2008). - PubMed
    1. Bhatia M., Bonnet D., Murdoch B., Gan O.I. & Dick J.E. A newly discovered class of human hematopoietic cells with SCID-repopulating activity. Nat. Med. 4, 1038–1045 (1998). - PubMed
    1. Scopelliti A. et al. Therapeutic implications of Cancer Initiating Cells. Expert Opin. Biol. Ther. 9, 1005–1016 (2009). - PubMed
    1. Li L. & Bhatia R. Stem cell quiescence. Clin. Cancer Res. 17, 4936–4941 (2011) - PMC - PubMed
    1. Hermann P.C., Bhaskar S., Cioffi M. & Heeschen C. Cancer stem cells in solid tumors. Semin. Cancer Biol. 20, 77–84 (2010). - PubMed

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