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Combination of 22-oxa-1,25-dihydroxyvitamin D3, a vitamin D3 derivative, with vitamin K2 (VK2) synergistically enhances cell differentiation but suppresses VK2-inducing apoptosis in HL-60 cells

Leukemia volume 16, pages 1519–1527 (2002)Cite this article

Abstract

We originally reported that vitamin K2 (VK2) effectively induces apoptosis in various types of primary cultured leukemia cells and leukemia cell lines in vitro. In addition, VK2 was shown to induce differentiation of leukemia cells when the cells were resistant against VK2-inducing apoptosis. A novel synthetic vitamin D3derivative, 22-oxa-1,25-dihydroxyvitamin D3 (OCT: oxacarcitriol) shows a more potent differentiation-inducing ability among myeloid leukemia cells in vitro with much lesser extent of the induction of hypercalcemia in vivo as compared to the effects of 1α,25(OH)2D3. In the present study, we focused on the effects of a combination of OCT plus VK2 on leukemia cells. Treatment of HL-60 cells with OCT for 72 h induces monocytic differentiation. A combination of OCT plus VK2 dramatically enhances monocytic differentiation as assessed by morphologic features, positivity for non-specific esterase staining, and cell surface antigen expressions. This combined effect far exceeds the maximum differentiation induction ability at the optimal concentrations of either OCT or VK2 alone. In addition, pronounced accumulation of the cells in the G0/G1 phase is observed by combined treatment with OCT plus VK2 as compared with each vitamin alone. In contrast to cell differentiation, caspase-3 activation and apoptosis induction in response to VK2 are significantly suppressed in the presence of OCT in HL-60 cells. These data suggest that monocytic differentiation and apoptosis induction of HL-60 cells are inversely regulated. Furthermore, pronounced induction of differentiation by combined treatment with VK2 plus OCT was also observed in four out of six cases of primary cultured acute myeloid leukemia cells in vitro, suggesting that VK2 plus OCT might be a potent combination for the differentiation-based therapy for acute myeloid leukemias.

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Acknowledgements

We thank Ayako Hirota for her excellent technical assistance. We also thank Eisai Co. (Tokyo, Japan) for providing VK2 (menaquinone-4), and Chugai Pharmaceutical Co. (Tokyo, Japan) for providing OCT and 1α,25(OH)2D3. This study was supported by a Grant-in-Aid for Science Research (C) from The Ministry of Education, Science, Sports and Culture of Japan to KM (No. 11671017) and a Health Science Research Grant from Research Committee for Idiopathic Hemopoietic Disorders of the Ministry of Health and Welfare in Japan to KO.

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  1. K Funato and K Miyazawa: KF and KM contributed equally to this report and should be regarded as co-first authors

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  1. First Department of Internal Medicine (Hematology/Oncology), Tokyo Medical University, Tokyo, Japan

    K Funato, K Miyazawa, M Yaguchi, A Gotoh & K Ohyashiki

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Funato, K., Miyazawa, K., Yaguchi, M. et al. Combination of 22-oxa-1,25-dihydroxyvitamin D3, a vitamin D3 derivative, with vitamin K2 (VK2) synergistically enhances cell differentiation but suppresses VK2-inducing apoptosis in HL-60 cells. Leukemia 16, 1519–1527 (2002). https://doi.org/10.1038/sj.leu.2402614

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