Abstract
Background
Human multiple myeloma (MM) remains an incurable hematological malignancy. We have reported that β-lapachone, a pure compound derived from a plant, can induce cell death in a variety of human carcinoma cells, including ovary, colon, lung, prostate, pancreas, and breast, suggesting a wide spectrum of anticancer activity.
Materials and Methods
We first studied anti-survival effects of β-lapachone in human MM cells by colony formation assay. To determine whether the differential inhibition of colony formation occurs through antiproliferative activity, we performed MTT assays. The cytotoxicity of β-lapachone on human peripheral blood mononuclear cells was also measured by MTT assay. To determine whether the cell death induced by β-lapachone occurs through necrosis or apoptosis, we used the propidium iodide staining procedure to determine the sub-G1 fraction, Annexin-V staining for externalization of phosphatidylserine, and fragmentation of cellular genomic DNA subjected to gel electrophoresis. To investigate the mechanism of anti-MM activity, we examined Bcl-2 expression, cytochrome C release, and poly (ADP ribose) polymerase cleavage by Western blot assay.
Results
We found that β-lapachone (less than 4 µM) inhibits cell survival and proliferation by triggering cell death with characteristics of apoptosis in ARH-77, HS Sultan, and MM.1S cell lines, in freshly derived patient MM cells (MM.As), MM cell lines resistant to dexamethasone (MM.1R), doxorubicin (DOX.40), mitoxantrone (MR.20), and mephalan (LR5). Importantly, after treatment with β-lapachone, we observed no apoptosis in peripheral blood mononuclear cells in either quiescent or proliferative states, freshly isolated from healthy donors. In β-lapachone treated ARH-77, cytochrome C was released from mitochondria to cytosol, and poly (ADP ribose) polymerase was cleaved, signature events of apoptosis. Finally, the apoptosis induced by β-lapachone in MM cells was not blocked by either interleukin-6 or Bcl-2, which confer multidrug resistance in MM.
Conclusions
Our results suggest potential therapeutic application of β-lapachone against MM, particularly to overcome drug resistance in relapsed patients.
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Acknowledgments
We thank Dr. Kenneth Anderson for providing MM cells and for critically reading the manuscript. This work was supported in part by Grant RO1 CA61253 from the National Institutes of Health.
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Li, Y., Li, C.J., Yu, D. et al. Potent Induction of Apoptosis by β-Lapachone in Human Multiple Myeloma Cell Lines and Patient Cells. Mol Med 6, 1008–1015 (2000). https://doi.org/10.1007/BF03402052
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DOI: https://doi.org/10.1007/BF03402052