Eradication of chemotherapy-resistant CD44+ human ovarian cancer stem cells in mice by intraperitoneal administration of clostridium perfringens enterotoxin.
Casagrande, Francesca MS 1; Cocco, Emiliano PhD 1; Bellone, Stefania PhD 1; Richter, Christine E. MD 1; Bellone, Marta MS 1; Todeschini, Paola MS 1; Siegel, Eric MS 2; Varughese, Joyce MD 1; Arin-Silasi, Dan MD 1; Azodi, Masoud MD 1; Rutherford, Thomas J. MD, PhD 1; Pecorelli, Sergio MD, PhD 3; Schwartz, Peter E. MD 1; Santin, Alessandro D. MD 1,*
[Article]
Cancer.
117(24):5519-5528, December 15, 2011.
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BACKGROUND: Emerging evidence has suggested that the capability to sustain tumor formation, growth, and chemotherapy resistance in ovarian as well as other human malignancies exclusively resides in a small proportion of tumor cells termed cancer stem cells. During the characterization of CD44 ovarian cancer stem cells, we found a high expression of the genes encoding for claudin-4. Because this tight junction protein is the natural high-affinity receptor for Clostridium perfringens enterotoxin (CPE), we have extensively investigated the sensitivity of ovarian cancer stem cells to CPE treatment in vitro and in vivo.
METHODS: Real-time polymerase chain reaction and flow cytometry were used to evaluate claudin-3/-4 expression in ovarian cancer stem cells. Small interfering RNA knockdown experiments and MTS assays were used to evaluate CPE-induced cytotoxicity against ovarian cancer stem cell lines in vitro. C.B-17/SCID mice harboring ovarian cancer stem cell xenografts were used to evaluate CPE therapeutic activity in vivo.
RESULTS: CD44 ovarian cancer stem cells expressed claudin-4 gene at significantly higher levels than matched autologous CD44- ovarian cancer cells, and regardless of their higher resistance to chemotherapeutic agents died within 1 hour after exposure to 1.0 [mu]g/mL of CPE in vitro. Conversely, small-interfering RNA-mediated knockdown of claudin-3/-4 expression in CD44 cancer stem cells significantly protected cancer stem cells from CPE-induced cytotoxicity. Importantly, multiple intraperitoneal administrations of sublethal doses of CPE in mice harboring xenografts of chemotherapy-resistant CD44 ovarian cancer stem cells had a significant inhibitory effect on tumor progression leading to the cure and/or long-term survival of all treated animals (ie, 100% reduction in tumor burden in 50% of treated mice; P < .0001).
CONCLUSIONS: CPE may represent an unconventional, potentially highly effective strategy to eradicate chemotherapy-resistant cancer stem cells. Cancer 2011;. (C) 2011 American Cancer Society.
Copyright (C) 2011 John Wiley & Sons, Inc.
