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Nemorosone blocks proliferation and induces apoptosis in leukemia cells.

Abstract

OBJECTIVE

This work is aimed at characterizing nemorosone, isolated from Clusia rosea, as a potential antileukemic agent. In addition, we analyzed its influence on hematopoiesis in a mouse model.

MATERIALS AND METHODS

The isolation of nemorosone was carried out employing the RP-HPLC (reversed phase high-performance liquid chromatography) technique. Cytotoxicity was assessed in human leukemia cell lines including parental and chemotherapy-refractory sublines based on the MTT compound. Its effects on the cell cycle were analyzed using FACS (fluorescence-activated cell sorting) and Western blot techniques. Studies on the drug-induced early apoptotic process were carried out by means of fluorescence microscopy. Major signal transducers and the enzymatic inhibition of immunoprecipitated Akt/PKB were detected by Western blot. Hematopoiesis was analyzed in NMRI nu/nu mice after chronic nemorosone treatment, measuring hematological parameters by conventional laboratory techniques.

RESULTS

Nemorosone proved cytotoxic in both parental and chemoresistant leukemia cell lines with IC50 values between 2.10 and 3.10 mg/ml. No cross-resistances could be detected. Cell cycle studies showed apoptosis induction accompanied by an increase in the G0/G1 population in both cell lines studied, whereas a significant decrease in the S-phase was found in Jurkat cells. Nemorosone induced a down-regulation of cyclins A, B1, D1, and E as well as a dephosphorylation of cdc2. Major signal transduction elements such as ERK1/2 and p38 MAPK, as well as important oncoproteins such as c-Myb and BCR/ABL were also found down-regulated. The enzymatic activity of immunoprecipitated Akt/PKB was substantially inhibited in vitro. Moreover, subchronic nemorosone treatment induced reversible monocytosis and thrombocytosis in the mouse model examined.

CONCLUSIONS

Here, we demonstrate for the first time that nemorosone exerts cytotoxicity in leukemia cells, partly by targeting the Akt/PKB signal transducer, affecting protein levels and cell cycle progression. Finally, in vivo studies suggest that nemorosone significantly affects hematopoiesis in mice.

Links

Authors+Show Affiliations

,

Abteilung für Klinische Pharmakologie, Ruhr-Universität Bochum, Germany. david.diaz-carballo@rub.de

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Source

MeSH

Animals
Antineoplastic Agents, Phytogenic
Apoptosis
Benzophenones
Cell Cycle
Cell Line, Tumor
Cell Proliferation
Clusia
Drug Delivery Systems
Drug Resistance, Neoplasm
Female
Hematopoiesis
Humans
Inhibitory Concentration 50
Leukemia
Mice
Mice, Nude
Proto-Oncogene Proteins c-akt
Signal Transduction

Pub Type(s)

Journal Article

Language

eng

PubMed ID

18793585

Citation

Díaz-Carballo, D, et al. "Nemorosone Blocks Proliferation and Induces Apoptosis in Leukemia Cells." International Journal of Clinical Pharmacology and Therapeutics, vol. 46, no. 8, 2008, pp. 428-39.
Díaz-Carballo D, Malak S, Freistühler M, et al. Nemorosone blocks proliferation and induces apoptosis in leukemia cells. Int J Clin Pharmacol Ther. 2008;46(8):428-39.
Díaz-Carballo, D., Malak, S., Freistühler, M., Elmaagacli, A., Bardenheuer, W., & Reusch, H. P. (2008). Nemorosone blocks proliferation and induces apoptosis in leukemia cells. International Journal of Clinical Pharmacology and Therapeutics, 46(8), pp. 428-39.
Díaz-Carballo D, et al. Nemorosone Blocks Proliferation and Induces Apoptosis in Leukemia Cells. Int J Clin Pharmacol Ther. 2008;46(8):428-39. PubMed PMID: 18793585.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Nemorosone blocks proliferation and induces apoptosis in leukemia cells. AU - Díaz-Carballo,D, AU - Malak,S, AU - Freistühler,M, AU - Elmaagacli,A, AU - Bardenheuer,W, AU - Reusch,H P, PY - 2008/9/17/pubmed PY - 2008/10/8/medline PY - 2008/9/17/entrez SP - 428 EP - 39 JF - International journal of clinical pharmacology and therapeutics JO - Int J Clin Pharmacol Ther VL - 46 IS - 8 N2 - OBJECTIVE: This work is aimed at characterizing nemorosone, isolated from Clusia rosea, as a potential antileukemic agent. In addition, we analyzed its influence on hematopoiesis in a mouse model. MATERIALS AND METHODS: The isolation of nemorosone was carried out employing the RP-HPLC (reversed phase high-performance liquid chromatography) technique. Cytotoxicity was assessed in human leukemia cell lines including parental and chemotherapy-refractory sublines based on the MTT compound. Its effects on the cell cycle were analyzed using FACS (fluorescence-activated cell sorting) and Western blot techniques. Studies on the drug-induced early apoptotic process were carried out by means of fluorescence microscopy. Major signal transducers and the enzymatic inhibition of immunoprecipitated Akt/PKB were detected by Western blot. Hematopoiesis was analyzed in NMRI nu/nu mice after chronic nemorosone treatment, measuring hematological parameters by conventional laboratory techniques. RESULTS: Nemorosone proved cytotoxic in both parental and chemoresistant leukemia cell lines with IC50 values between 2.10 and 3.10 mg/ml. No cross-resistances could be detected. Cell cycle studies showed apoptosis induction accompanied by an increase in the G0/G1 population in both cell lines studied, whereas a significant decrease in the S-phase was found in Jurkat cells. Nemorosone induced a down-regulation of cyclins A, B1, D1, and E as well as a dephosphorylation of cdc2. Major signal transduction elements such as ERK1/2 and p38 MAPK, as well as important oncoproteins such as c-Myb and BCR/ABL were also found down-regulated. The enzymatic activity of immunoprecipitated Akt/PKB was substantially inhibited in vitro. Moreover, subchronic nemorosone treatment induced reversible monocytosis and thrombocytosis in the mouse model examined. CONCLUSIONS: Here, we demonstrate for the first time that nemorosone exerts cytotoxicity in leukemia cells, partly by targeting the Akt/PKB signal transducer, affecting protein levels and cell cycle progression. Finally, in vivo studies suggest that nemorosone significantly affects hematopoiesis in mice. SN - 0946-1965 UR - https://www.unboundmedicine.com/medline/citation/18793585/Nemorosone_blocks_proliferation_and_induces_apoptosis_in_leukemia_cells_ L2 - https://medlineplus.gov/leukemia.html DB - PRIME DP - Unbound Medicine ER -