Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization

Abstract Background Gynostemma pentaphyllum (Thunb.), a traditional adaptogenic herb, is known for its bioactive components with potential anti-cancer properties. Acute myeloid leukemia (AML) progression is significantly influenced by Feline McDonough Sarcoma (FMS)-like tyrosine kinase 3 (FLT3) sign...

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Main Authors: Khin Khin Gyi, Songyot Anuchapreeda, Nutjeera Intasai, Montree Tungjai, Siriporn Okonogi, Arihiro Iwasaki, Toyonobu Usuki, Singkome Tima
Format: Article
Language:English
Published: BMC 2025-05-01
Series:BMC Complementary Medicine and Therapies
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Online Access:https://doi.org/10.1186/s12906-025-04903-0
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author Khin Khin Gyi
Songyot Anuchapreeda
Nutjeera Intasai
Montree Tungjai
Siriporn Okonogi
Arihiro Iwasaki
Toyonobu Usuki
Singkome Tima
author_facet Khin Khin Gyi
Songyot Anuchapreeda
Nutjeera Intasai
Montree Tungjai
Siriporn Okonogi
Arihiro Iwasaki
Toyonobu Usuki
Singkome Tima
author_sort Khin Khin Gyi
collection DOAJ
description Abstract Background Gynostemma pentaphyllum (Thunb.), a traditional adaptogenic herb, is known for its bioactive components with potential anti-cancer properties. Acute myeloid leukemia (AML) progression is significantly influenced by Feline McDonough Sarcoma (FMS)-like tyrosine kinase 3 (FLT3) signaling, while Wilms’ tumor 1 (WT1) serves as a key prognostic marker. This study investigates the anti-leukemia activities of active G. pentaphyllum leaf extracts and their components, focusing on the inhibition of FLT3 and WT1 activity. Methods G. pentaphyllum extracts were prepared through maceration, yielding three crude fractional extracts. The cytotoxicity of the extracts was screened against various leukemia cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The most cytotoxic extract was further fractionated and purified via column chromatography. The anti-proliferative and apoptotic induction activities of the active extract and its fraction were evaluated through cell cycle and apoptosis analyses using flow cytometry. Changes in mitochondrial membrane potential (ΔΨm) were assessed by spectrofluorometry. To confirm anti-leukemia activity, the expression levels of FLT3, WT1 and apoptotic-related protein were analyzed using Western blotting. The major active compounds within the active fractions were identified and characterized using Electrospray Ionization Mass Spectrometry (ESI-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy. Results The ethyl acetate fractional extract (F-EtOAc) demonstrated the highest cytotoxicity, particularly against FLT3-overexpressing EoL-1 (IC50 = 40.82 ± 0.8 µg/mL) and MV4-11 (IC50 = 35.54 ± 4.1 µg/mL) AML cell lines. Fraction F10 was identified as the most active fraction, significantly inhibited FLT3 and WT1 protein expression and induced G0/G1 cell cycle arrest in a dose-dependent manner. Additionally, F10 induced dose-dependent apoptosis through disruption of ΔΨm, p53 up-regulation and caspase-3 activation. Further purification of F10 identified dehydrovomifoliol as its major bioactive compound. Conclusion These findings suggest that the ethyl acetate extract of G. pentaphyllum contains bioactive compounds with anti-leukemia potential, warranting further investigation to evaluate its efficacy against AML. Clinical trial number Not applicable.
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spelling doaj-art-d57a2aa9ccd94624abbf80bc630839b82025-08-20T03:10:16ZengBMCBMC Complementary Medicine and Therapies2662-76712025-05-0125111810.1186/s12906-025-04903-0Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterizationKhin Khin Gyi0Songyot Anuchapreeda1Nutjeera Intasai2Montree Tungjai3Siriporn Okonogi4Arihiro Iwasaki5Toyonobu Usuki6Singkome Tima7Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityDepartment of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityDepartment of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityDepartment of Radiologic Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityCenter of Excellence in Pharmaceutical Nanotechnology, Chiang Mai UniversityDepartment of Applied Chemistry, Faculty of Science and Engineering, Chuo UniversityDepartment of Materials and Life Sciences, Faculty of Science and Technology, Sophia UniversityDepartment of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai UniversityAbstract Background Gynostemma pentaphyllum (Thunb.), a traditional adaptogenic herb, is known for its bioactive components with potential anti-cancer properties. Acute myeloid leukemia (AML) progression is significantly influenced by Feline McDonough Sarcoma (FMS)-like tyrosine kinase 3 (FLT3) signaling, while Wilms’ tumor 1 (WT1) serves as a key prognostic marker. This study investigates the anti-leukemia activities of active G. pentaphyllum leaf extracts and their components, focusing on the inhibition of FLT3 and WT1 activity. Methods G. pentaphyllum extracts were prepared through maceration, yielding three crude fractional extracts. The cytotoxicity of the extracts was screened against various leukemia cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The most cytotoxic extract was further fractionated and purified via column chromatography. The anti-proliferative and apoptotic induction activities of the active extract and its fraction were evaluated through cell cycle and apoptosis analyses using flow cytometry. Changes in mitochondrial membrane potential (ΔΨm) were assessed by spectrofluorometry. To confirm anti-leukemia activity, the expression levels of FLT3, WT1 and apoptotic-related protein were analyzed using Western blotting. The major active compounds within the active fractions were identified and characterized using Electrospray Ionization Mass Spectrometry (ESI-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy. Results The ethyl acetate fractional extract (F-EtOAc) demonstrated the highest cytotoxicity, particularly against FLT3-overexpressing EoL-1 (IC50 = 40.82 ± 0.8 µg/mL) and MV4-11 (IC50 = 35.54 ± 4.1 µg/mL) AML cell lines. Fraction F10 was identified as the most active fraction, significantly inhibited FLT3 and WT1 protein expression and induced G0/G1 cell cycle arrest in a dose-dependent manner. Additionally, F10 induced dose-dependent apoptosis through disruption of ΔΨm, p53 up-regulation and caspase-3 activation. Further purification of F10 identified dehydrovomifoliol as its major bioactive compound. Conclusion These findings suggest that the ethyl acetate extract of G. pentaphyllum contains bioactive compounds with anti-leukemia potential, warranting further investigation to evaluate its efficacy against AML. Clinical trial number Not applicable.https://doi.org/10.1186/s12906-025-04903-0Acute myeloid leukemiaGynostemma pentaphyllum (Thunb.)FLT3WT1Anti-leukemiaDehydrovomifoliol
spellingShingle Khin Khin Gyi
Songyot Anuchapreeda
Nutjeera Intasai
Montree Tungjai
Siriporn Okonogi
Arihiro Iwasaki
Toyonobu Usuki
Singkome Tima
Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization
BMC Complementary Medicine and Therapies
Acute myeloid leukemia
Gynostemma pentaphyllum (Thunb.)
FLT3
WT1
Anti-leukemia
Dehydrovomifoliol
title Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization
title_full Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization
title_fullStr Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization
title_full_unstemmed Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization
title_short Anti-leukemia activity of the ethyl acetate extract from Gynostemma pentaphyllum (Thunb.) leaf against FLT3-overexpressing AML cells and its phytochemical characterization
title_sort anti leukemia activity of the ethyl acetate extract from gynostemma pentaphyllum thunb leaf against flt3 overexpressing aml cells and its phytochemical characterization
topic Acute myeloid leukemia
Gynostemma pentaphyllum (Thunb.)
FLT3
WT1
Anti-leukemia
Dehydrovomifoliol
url https://doi.org/10.1186/s12906-025-04903-0
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