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19-Nor-1α,25-dihydroxyvitamin D2 (paricalcitol): effects on clonal proliferation, differentiation, and apoptosis in human leukemic cell lines

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Abstract

Purpose

19-Nor-1α,25-dihydroxyvitamin D2 (paricalcitol) is an analogue of 1,25(OH)2D3 with reduced calcemic effects that is approved for the suppression of parathyroid hormone in chronic renal failure. Paricalcitol has recently been reported to have anticancer activity in prostate cancer. In order to explore paricalcitol as a potential agent against leukemia, we tested its effects on HL-60 and U937 leukemia cell lines.

Methods

We studied cellular differentiation via expression of CD11b and CD14 surface antigens using flow cytometry, and via the nitroblue tetrazolium (NBT) assay. Cell cycle was analyzed using propidium iodide staining. Apoptosis was assessed with the annexin V assay. Cellular proliferation was determined via colony inhibition on semisolid medium.

Results

Paricalcitol induced the maturation of HL-60 and U937 cells, as shown by increased expression of CD11b differentiation surface antigen. CD14 showed increased expression in HL-60 but not in U937 cells. After exposure to paricalcitol at 10−8  M for 72 h, the ability of HL-60 cells to reduce NBT was markedly increased. Conversely, U937 cells were unchanged. Paricalcitol inhibited colony formation of both HL-60 and U937 cell lines in semisolid medium after a 10-day incubation (estimated IC50 of 3×10−8  M in HL-60 cells and 4×10−8 M in U937 cells). Paricalcitol at 10−8  M and 10−7  M caused a significant dose- and time-dependent increase of apoptosis in HL-60 cells (P<0.05). In both HL-60 and U937 cells, exposure to 10−7  M paricalcitol for 72 h increased the number of cells in G0/G1 phase, and decreased the number of cells in S phase.

Conclusions

Paricalcitol inhibits colony formation, induces maturation and causes cell cycle arrest in HL-60 and U937 cells. Additionally, paricalcitol induces apoptosis in HL-60 cells. These findings support the further evaluation of paricalcitol as an antileukemia agent.

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Acknowledgements

This research was supported, in part, by developmental funds (PUSH grant) from the Comprehensive Cancer Center of Wake Forest University. Paricalcitol was a generous gift from Abbott Laboratories, Chicago, Ill.

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Authors and Affiliations

  1. Department of Internal Medicine, Section on Hematology and Oncology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA

    István Molnár & Bayard L. Powell

  2. Department of Pathology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA

    Timothy Kute, Mark C. Willingham & William H. Dodge

  3. Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA

    Gary G. Schwartz

  4. Comprehensive Cancer Center, Wake Forest University, Medical Center Blvd, Winston-Salem, NC 27157, USA

    Gary G. Schwartz

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  1. István Molnár
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  2. Timothy Kute
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  3. Mark C. Willingham
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  4. Bayard L. Powell
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  5. William H. Dodge
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  6. Gary G. Schwartz
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Correspondence to Gary G. Schwartz.

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Molnár, I., Kute, T., Willingham, M.C. et al. 19-Nor-1α,25-dihydroxyvitamin D2 (paricalcitol): effects on clonal proliferation, differentiation, and apoptosis in human leukemic cell lines. J Cancer Res Clin Oncol 129, 35–42 (2003). https://doi.org/10.1007/s00432-002-0405-7

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  • DOI: https://doi.org/10.1007/s00432-002-0405-7

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