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2-Hydroxypropyl-β-Cyclodextrin Acts as a Novel Anticancer Agent.

Yokoo M, Kubota Y, Motoyama K, Higashi T, Taniyoshi M, Tokumaru H, Nishiyama R, Tabe Y, Mochinaga S, Sato A, Sueoka-Aragane N, Sueoka E, Arima H, Irie T, Kimura S - PLoS ONE (2015)

Bottom Line: HP-β-CyD treatment reduced intracellular cholesterol resulting in significant leukemic cell growth inhibition through G2/M cell-cycle arrest and apoptosis.Importantly, HP-β-CyD also showed anticancer effects against CML cells expressing a T315I BCR-ABL mutation (that confers resistance to most ABL tyrosine kinase inhibitors), and hypoxia-adapted CML cells that have characteristics of leukemic stem cells.Systemic administration of HP-β-CyD to mice had no significant adverse effects.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology, Respiratory Medicine and Oncology, Department of Internal Medicine, Faculty of Medicine, Saga University, Saga, Japan.

ABSTRACT
2-Hydroxypropyl-β-cyclodextrin (HP-β-CyD) is a cyclic oligosaccharide that is widely used as an enabling excipient in pharmaceutical formulations, but also as a cholesterol modifier. HP-β-CyD has recently been approved for the treatment of Niemann-Pick Type C disease, a lysosomal lipid storage disorder, and is used in clinical practice. Since cholesterol accumulation and/or dysregulated cholesterol metabolism has been described in various malignancies, including leukemia, we hypothesized that HP-β-CyD itself might have anticancer effects. This study provides evidence that HP-β-CyD inhibits leukemic cell proliferation at physiologically available doses. First, we identified the potency of HP-β-CyD in vitro against various leukemic cell lines derived from acute myeloid leukemia (AML), acute lymphoblastic leukemia and chronic myeloid leukemia (CML). HP-β-CyD treatment reduced intracellular cholesterol resulting in significant leukemic cell growth inhibition through G2/M cell-cycle arrest and apoptosis. Intraperitoneal injection of HP-β-CyD significantly improved survival in leukemia mouse models. Importantly, HP-β-CyD also showed anticancer effects against CML cells expressing a T315I BCR-ABL mutation (that confers resistance to most ABL tyrosine kinase inhibitors), and hypoxia-adapted CML cells that have characteristics of leukemic stem cells. In addition, colony forming ability of human primary AML and CML cells was inhibited by HP-β-CyD. Systemic administration of HP-β-CyD to mice had no significant adverse effects. These data suggest that HP-β-CyD is a promising anticancer agent regardless of disease or cellular characteristics.

No MeSH data available.


Related in: MedlinePlus

HP-β-CyD causes cell-cycle arrest in leukemic cells.(A–D) BV173 and K562 cells were treated with the indicated concentration of HP-β-CyD for 12 hours, then flow cytometric analysis of PI-stained nuclei was performed. (A) Representative flow cytometric histograms of PI-stained BV173 cells. (B) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable BV173 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (C) Representative flow cytometric histograms of PI-stained K562 cells. (D) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable K562 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (E and F) Effects of HP-β-CyD on the expression of G2/M cell-cycle-associated proteins. BV173 (E) and K562 cells (F) were treated with 10 mM HP-β-CyD for the indicated times. Cells were lysed and analyzed by Western blotting. Western blot images are representative results from at least two independent experiments. Detection of β-actin was used as a loading control. Intensity of the immunoblot signals after background subtraction was quantified using ImageJ software.
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pone.0141946.g003: HP-β-CyD causes cell-cycle arrest in leukemic cells.(A–D) BV173 and K562 cells were treated with the indicated concentration of HP-β-CyD for 12 hours, then flow cytometric analysis of PI-stained nuclei was performed. (A) Representative flow cytometric histograms of PI-stained BV173 cells. (B) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable BV173 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (C) Representative flow cytometric histograms of PI-stained K562 cells. (D) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable K562 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (E and F) Effects of HP-β-CyD on the expression of G2/M cell-cycle-associated proteins. BV173 (E) and K562 cells (F) were treated with 10 mM HP-β-CyD for the indicated times. Cells were lysed and analyzed by Western blotting. Western blot images are representative results from at least two independent experiments. Detection of β-actin was used as a loading control. Intensity of the immunoblot signals after background subtraction was quantified using ImageJ software.

Mentions: Furthermore, both BV173 (Fig 3A and 3B) and K562 cells (Fig 3C and 3D) responded to HP-β-CyD with a dose-dependent increase in the percentage of cells in G2/M phase. As the percentage of cells in G2/M increased, the percentage of G1 phase decreased, whereas the proportion of S-phase cells was not significantly altered by HP-β-CyD treatment. Treatment with HP-β-CyD also induced G2/M cell-cycle arrest in another Ph+ cell line, KBM5, and Ph-negative cell lines such as NALM-6, Jurkat, and MOLT4 (S1 Fig). Next, the effects of HP-β-CyD treatment on the expression of G2/M cell-cycle regulators, such as cdc25C, cdc2, cyclin A, cyclin B1, and p21, were examined by western blot analysis. HP-β-CyD had no effect on p-cdc25c and cyclin B1 protein expression; however, levels of cdc2 slightly decreased in BV173 cells (Fig 3E and 3F). Cyclin A and p21 decreased in BV173 cells, while in K562 cells, p21 expression was induced after 6 hours of HP-β-CyD treatment and slowly declined thereafter (Fig 3E and 3F). p21 is a cell cycle regulatory protein that can cause cell cycle arrest [41]. However, the role of p21 is complex because it also posseses pro- and anti-apoptotic abilities, and the function of p21 in apoptosis is cell type and cellular context specific [42–46]. Therefore, different behavior of p21 in HP-β-CyD-treated BV173 and K562 cells may be explained in part by the multiple roles of p21. These data indicate that HP-β-CyD-induced cell growth inhibition could be, in part, ascribed to induction of apoptosis and cell-cycle arrest at the G2/M checkpoint.


2-Hydroxypropyl-β-Cyclodextrin Acts as a Novel Anticancer Agent.

Yokoo M, Kubota Y, Motoyama K, Higashi T, Taniyoshi M, Tokumaru H, Nishiyama R, Tabe Y, Mochinaga S, Sato A, Sueoka-Aragane N, Sueoka E, Arima H, Irie T, Kimura S - PLoS ONE (2015)

HP-β-CyD causes cell-cycle arrest in leukemic cells.(A–D) BV173 and K562 cells were treated with the indicated concentration of HP-β-CyD for 12 hours, then flow cytometric analysis of PI-stained nuclei was performed. (A) Representative flow cytometric histograms of PI-stained BV173 cells. (B) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable BV173 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (C) Representative flow cytometric histograms of PI-stained K562 cells. (D) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable K562 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (E and F) Effects of HP-β-CyD on the expression of G2/M cell-cycle-associated proteins. BV173 (E) and K562 cells (F) were treated with 10 mM HP-β-CyD for the indicated times. Cells were lysed and analyzed by Western blotting. Western blot images are representative results from at least two independent experiments. Detection of β-actin was used as a loading control. Intensity of the immunoblot signals after background subtraction was quantified using ImageJ software.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4633159&req=5

pone.0141946.g003: HP-β-CyD causes cell-cycle arrest in leukemic cells.(A–D) BV173 and K562 cells were treated with the indicated concentration of HP-β-CyD for 12 hours, then flow cytometric analysis of PI-stained nuclei was performed. (A) Representative flow cytometric histograms of PI-stained BV173 cells. (B) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable BV173 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (C) Representative flow cytometric histograms of PI-stained K562 cells. (D) The percentage of cells in G0/G1, S, or G2/M phase was assessed in viable K562 cells. White: G1-phase, gray: S-phase, black: G2/M-phase. (E and F) Effects of HP-β-CyD on the expression of G2/M cell-cycle-associated proteins. BV173 (E) and K562 cells (F) were treated with 10 mM HP-β-CyD for the indicated times. Cells were lysed and analyzed by Western blotting. Western blot images are representative results from at least two independent experiments. Detection of β-actin was used as a loading control. Intensity of the immunoblot signals after background subtraction was quantified using ImageJ software.
Mentions: Furthermore, both BV173 (Fig 3A and 3B) and K562 cells (Fig 3C and 3D) responded to HP-β-CyD with a dose-dependent increase in the percentage of cells in G2/M phase. As the percentage of cells in G2/M increased, the percentage of G1 phase decreased, whereas the proportion of S-phase cells was not significantly altered by HP-β-CyD treatment. Treatment with HP-β-CyD also induced G2/M cell-cycle arrest in another Ph+ cell line, KBM5, and Ph-negative cell lines such as NALM-6, Jurkat, and MOLT4 (S1 Fig). Next, the effects of HP-β-CyD treatment on the expression of G2/M cell-cycle regulators, such as cdc25C, cdc2, cyclin A, cyclin B1, and p21, were examined by western blot analysis. HP-β-CyD had no effect on p-cdc25c and cyclin B1 protein expression; however, levels of cdc2 slightly decreased in BV173 cells (Fig 3E and 3F). Cyclin A and p21 decreased in BV173 cells, while in K562 cells, p21 expression was induced after 6 hours of HP-β-CyD treatment and slowly declined thereafter (Fig 3E and 3F). p21 is a cell cycle regulatory protein that can cause cell cycle arrest [41]. However, the role of p21 is complex because it also posseses pro- and anti-apoptotic abilities, and the function of p21 in apoptosis is cell type and cellular context specific [42–46]. Therefore, different behavior of p21 in HP-β-CyD-treated BV173 and K562 cells may be explained in part by the multiple roles of p21. These data indicate that HP-β-CyD-induced cell growth inhibition could be, in part, ascribed to induction of apoptosis and cell-cycle arrest at the G2/M checkpoint.

Bottom Line: HP-β-CyD treatment reduced intracellular cholesterol resulting in significant leukemic cell growth inhibition through G2/M cell-cycle arrest and apoptosis.Importantly, HP-β-CyD also showed anticancer effects against CML cells expressing a T315I BCR-ABL mutation (that confers resistance to most ABL tyrosine kinase inhibitors), and hypoxia-adapted CML cells that have characteristics of leukemic stem cells.Systemic administration of HP-β-CyD to mice had no significant adverse effects.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology, Respiratory Medicine and Oncology, Department of Internal Medicine, Faculty of Medicine, Saga University, Saga, Japan.

ABSTRACT
2-Hydroxypropyl-β-cyclodextrin (HP-β-CyD) is a cyclic oligosaccharide that is widely used as an enabling excipient in pharmaceutical formulations, but also as a cholesterol modifier. HP-β-CyD has recently been approved for the treatment of Niemann-Pick Type C disease, a lysosomal lipid storage disorder, and is used in clinical practice. Since cholesterol accumulation and/or dysregulated cholesterol metabolism has been described in various malignancies, including leukemia, we hypothesized that HP-β-CyD itself might have anticancer effects. This study provides evidence that HP-β-CyD inhibits leukemic cell proliferation at physiologically available doses. First, we identified the potency of HP-β-CyD in vitro against various leukemic cell lines derived from acute myeloid leukemia (AML), acute lymphoblastic leukemia and chronic myeloid leukemia (CML). HP-β-CyD treatment reduced intracellular cholesterol resulting in significant leukemic cell growth inhibition through G2/M cell-cycle arrest and apoptosis. Intraperitoneal injection of HP-β-CyD significantly improved survival in leukemia mouse models. Importantly, HP-β-CyD also showed anticancer effects against CML cells expressing a T315I BCR-ABL mutation (that confers resistance to most ABL tyrosine kinase inhibitors), and hypoxia-adapted CML cells that have characteristics of leukemic stem cells. In addition, colony forming ability of human primary AML and CML cells was inhibited by HP-β-CyD. Systemic administration of HP-β-CyD to mice had no significant adverse effects. These data suggest that HP-β-CyD is a promising anticancer agent regardless of disease or cellular characteristics.

No MeSH data available.


Related in: MedlinePlus