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

Pediatric Surgical Research Laboratories, Department of Surgery, Harvard Medical School, 185 Cambridge Street, Boston, MA 02114, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 08/2006; 103(30):11154-9. DOI: 10.1073/pnas.0603672103
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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.

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Available from: Rosemary Foster, Jan 05, 2015
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    • "We and others have recently demonstrated an association between chemoresistance and CSC-like phenotypes in ovarian cancer [9–12], and found chemoresistant recurrent ovarian tumors to be enriched in CSCs and stem cell pathway mediators, suggesting that CSCs may contribute to recurrent disease [13, 14]. CSCs have also been isolated from ovarian cancer cell lines based on their abilities to differentially efflux the DNA binding dye Hoechst 33342 [15]. This population of cells termed the 'side population' (SP) displayed the classical stem cell property in tumorigenicity assays. "
    02/2015; 3(1):3:e1001. DOI:10.14343/JCSCR.2015.3e1001
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    • "We next analyzed the proportion of the SP fraction in shCHIP cells to further assess whether CHIP suppressed CSC phenotypes. SP assay is one method used to assess CSC enrichment in several cancers, including breast cancer [25], hepatocellular carcinoma [26], ovarian cancer [27], and lung cancer [28]. SP cells, which have the ability to efflux the DNA-binding dye Hoechst 33342 out of the cell membranes, were detected using FACS analysis [20]. "
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    ABSTRACT: Cancer stem cells (CSCs) have several distinctive characteristics, including high metastatic potential, tumor-initiating potential, and properties that resemble normal stem cells such as self-renewal, differentiation, and drug efflux. Because of these characteristics, CSC is regarded to be responsible for cancer progression and patient prognosis. In our previous study, we showed that a ubiquitin E3 ligase carboxyl terminus of Hsc70-interacting protein (CHIP) suppressed breast cancer malignancy. Moreover, a recent clinical study reported that CHIP expression levels were associated with favorable prognostic parameters of patients with breast cancer. Here we show that CHIP suppresses CSC properties in a population of breast cancer cells. CHIP depletion resulted in an increased proportion of CSCs among breast cancers when using several assays to assess CSC properties. From our results, we propose that inhibition of CSC properties may be one of the functions of CHIP as a suppressor of cancer progression.
    Biochemical and Biophysical Research Communications 09/2014; 452(4). DOI:10.1016/j.bbrc.2014.09.011 · 2.30 Impact Factor
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    • "493 confirmed using in vitro tumorsphere assays and in vivo animal assays (data not shown). There are many well-established methods that demonstrate the isolation of CSCs from different cancers using Hoechst 33342 dye and surface markers (Al-Hajj et al, 2003; Collins et al, 2005; Dean et al, 2005; Szotek et al, 2006; Dalerba et al, 2007; Mimeault et al, 2007; Engelmann et al, 2008; Ferrandina et al, 2008; Ponnusamy and Batra, 2008; Marsden et al, 2009). The main limitation of using Hoechst dye is its toxicity to cells; but if the concentration and incubation time has been standardised toxicity to cells could be minimised. "
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    ABSTRACT: Background: Cancer stem cells (CSCs) contribute towards disease aggressiveness and drug resistance. Specific identification of CSC maintenance genes and targeting can improve the efficiency of currently available treatment modalities. Pancreatic differentiation 2 (PD2) has a major role in the self-renewal of mouse embryonic stem cells. In the present study, we investigated the role of PD2 in pancreatic CSCs. Methods: Characterisation of CSCs and non-CSCs from mouse models, pancreatic cancer cells and human tissues by CSC and self-renewal marker analysis using confocal assay. Effect of PD2 knockdown in CSCs (after gemcitabine treatment) was studied by immunoblot and apoptosis assays. Results: A subpopulation of cells displayed PD2 overexpression in mouse (KrasG12D; Pdx1-Cre and KrasG12D; Trp53R172H/+; Pdx1-Cre) and human pancreatic tumours, which co-express CSC markers. Cancer stem cells exhibited elevated expression of PD2 and self-renewal markers, such as Oct3/4, Shh and β-catenin. Gemcitabine treatment maintained the CSC population with simultaneous maintenance of PD2 and CSC marker expression. Knockdown of PD2 in CSCs resulted in reduced viability of cells and enhanced apoptosis along with abrogated expression of CD133 and MDR2. Conclusions: Our results suggest that PD2 is a novel CSC maintenance protein, loss of which renders the CSCs more susceptible to drug-induced cell death.
    British Journal of Cancer 07/2014; DOI:10.1038/bjc.2014.152 · 4.84 Impact Factor
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