Abstract
Purpose
TRAIL/Apo-2L is cytotoxic against numerous prostate tumor cell lines; however, some lines are more resistant than others. Identification of an agent that increases prostate tumor cell sensitivity to TRAIL/Apo-2L would prove valuable for TRAIL/Apo-2L-mediated tumor therapy. Thus, we examined the effect of combining five clinically approved chemotherapeutic agents with TRAIL/Apo-2L for treating prostate tumor cells.
Methods
Four human prostate tumor cell lines were initially tested for TRAIL/Apo-2L sensitivity. Subsequent studies examined whether the TRAIL/Apo-2L-induced killing of DU-145 cells was augmented in the presence of the chemotherapeutic molecules, as measured by annexin V-FITC/propidium iodide staining. Furthermore, caspase 8 activation and BID cleavage were examined by immunoblotting. RT-PCR and flow cytometry were performed to monitor TRAIL-R1 and TRAIL-R2 levels after chemotherapeutic treatment.
Results
DU-145 cells were the least responsive of the prostate tumor cell lines tested to TRAIL/Apo-2L-induced death. Surprisingly, only topotecan, a topoisomerase I inhibitor, when used in combination with rTRAIL/Apo-2L led to significant apoptosis of DU-145 cells, as measured by caspase 8 activation, BID cleavage, and annexin V-FITC/PI staining. Topotecan alone had little to no toxicity on the DU-145 cells. Furthermore, the increase in TRAIL/Apo-2L sensitivity following topotecan treatment correlated with increased expression of TRAIL-R1 and TRAIL-R2 and decreased intracellular levels of the antiapoptotic protein survivin.
Conclusions
Our results define a promising direction for alternative therapies against androgen-independent prostate cancers. The sensitivity of DU-145 cells to TRAIL/Apo-2L was dramatically increased when combined with topotecan, suggesting that low-dose topotecan treatment to upregulate TRAIL-R1 and TRAIL-R2 and downregulate survivin, followed by TRAIL/Apo-2L administration, may be a viable therapy for treating cancer of the prostate.
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Abbreviations
- BID:
-
Bcl-2-interacting domain death agonist
- FADD:
-
Fas-associated death domain
- FLIP:
-
FADD-like IL-1β-converting enzyme-inhibitory protein
- IAP:
-
Inhibitor of apoptosis protein
- MDR:
-
Multidrug resistance
- PrEC:
-
Prostate epithelial cells
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- TRAIL:
-
TNF-related apoptosis-inducing ligand
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Acknowledgements
We thank Drs. Ben Elzey and Tim Ratliff for careful reading the manuscript. We would also like to acknowledge Linda Buckner for secretarial assistance. This work was supported by an American Cancer Society Institutional Seed Grant (no. IN-122U) administered through the University of Iowa's Holden Comprehensive Cancer Center, the Edwin Beer Program of The New York Academy of Medicine, and a Department of Defense Prostate Cancer Research Program New Investigator Award (PC010599).
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Griffith, T.S., Kemp, T.J. The topoisomerase I inhibitor topotecan increases the sensitivity of prostate tumor cells to TRAIL/Apo-2L-induced apoptosis. Cancer Chemother Pharmacol 52, 175–184 (2003). https://doi.org/10.1007/s00280-003-0656-2
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DOI: https://doi.org/10.1007/s00280-003-0656-2