Elucidating the inhibitory mechanisms of the ethanolic extract of the fruiting body of the mushroom Antrodia cinnamomea on the proliferation and migration of murine leukemia WEHI-3 cells and their tumorigenicity in a BALB/c allograft tumor model
Graphical abstract
Introduction
The survival of patients with acute myeloid leukemia (AML) is poor, with only 21.4% and 18.7% of patients surviving for 5 and 10 years after diagnosis, respectively (Pulte et al. 2010). In clinical trials, arsenic trioxide (ATO) and all-trans retinoic acid (ATRA) have been shown to be efficacious in the treatment of patients with both newly diagnosed and relapsing acute promyelocytic leukemia (APL), a subtype of AML, and standard AML (Amadori et al. 2005). However, in clinical trials, single agent treatment with ATO has not demonstrated any significant benefits in a variety of non-APL hematological malignancies (Lu et al. 2012b). Additionally long-term treatment with ATRA and ATO can result in several serious side effects, including hypertension (Li et al. 2010), interstitial pulmonary infiltrates, pleural and pericardial effusion, dyspnea, episodic hypotension and acute renal failure (Patatanian and Thompson 2008). Thus, new therapeutic agents are needed for the treatment of AML.
It is generally believed that the proliferation and migration of leukemia cells are involved in tumor growth and invasion. Therefore, modulation of the growth and migration properties of leukemia cells has important therapeutic implications. To explore this possible approach, we investigated the ability of Antrodia cinnamomea (AC), a medical mushroom from Taiwan, to modulate the proliferation and migration of leukemia cells in vitro, as well as the antitumor effects of AC in vivo.
Traditionally, AC has been used as a health food to prevent inflammation, hypertension, itchy skin, and liver cancer (Liaw 1985). There is increasing evidence that AC possesses an extensive range of biological properties, including protection against hepatotoxicity (Ao et al. 2009), anti-inflammatory properties (Chen et al. 2007), and antioxidant properties (Yang et al. 2006). Several studies have suggested that the extracts from the mycelia and the fruiting bodies of AC could be used as potential chemotherapeutic agents against various cancers, including hepatoma, prostate, colon, breast, bladder, and lung cancer (Hseu et al., 2008, Lin et al., 2010, Wu et al., 2006, Yang et al., 2011, Yeh et al., 2009). Our previous studies have identified that three main constituents of the EEAC, including adenosine, cordycepin and zhankuic acid A in a HPLC/Mass-fingerprint analysis (Chen et al. 2012b). However, the antimetastatic and antitumor effects of the fruiting bodies of AC in acute myeloid leukemia are still unclear. The present study was designed to explore the mechanisms of the antimetastatic effects of the ethanolic extract of AC (EEAC), cordycepin and zhankuic acid A on the WEHI-3 cell line, as well as the antitumor effects of EEAC in BALB/c mice injected with WEHI-3 cells.
Section snippets
Materials
Penicillin G, M199 (31100-035), FBS (10099-141) medium and streptomycin were obtained from Invitrogen (Carlsbad, CA, USA). Primary antibodies against phosphorylated-ERK1/2(p-ERK1/2, #sc-7383) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against phosphorylated-Akt (p-Akt, #ab28821), MMP-9 (#ab58803), p21 (#ab7960) and β-actin (#ab8226) were purchased from Abcam (Cambridge, MA, USA). Antibodies against p27 (GTX100446), Cyclin D1 (GTX27958) and Cyclin E (GTX27959)
The effect of EEAC extract on the proliferation of WEHI-3 leukemic cells
The anti-proliferative effect of EEAC was analyzed using WEHI-3 cells, which were treated with 10% FBS to stimulate cell growth. The treatment of WEHI-3 cells with 6.25 to 25 μg/ml EEAE for 24 h significantly suppressed their proliferation, with a 50% inhibitory concentration (IC50) of 15 μg/ml (Fig. 1A). In contrast, HUVEC showed decreased proliferation only when incubated with EEAC at 40–60 μg/ml for 24 h, and HUVEC cell viability was not significantly affected by lower concentrations of EEAC,
Discussion
Abnormal cell proliferation and metastasis play critical biological roles in clinical cancer cases. Metastasis, the invasion of cancer cells into other organs, is a complicated process and is an important and characteristic step in cancer progression, resulting in poor clinical outcome. As such, a candidate drug that efficiently inhibits tumor growth and the invasion and migration of cancer cells may suppress cancer progression and metastasis, potentially even reducing mortality. In the present
Conflict of interest
All authors reviewed the manuscript, agreed with its contents, declared that they do not have any conflicts of interest in regard to the subject matter or materials in the research and approved its submission for publication.
Acknowledgements
The authors wish to thank Yu-Hsuan Lin, Tai-Yun Wu and Pei-Hua Tu for assistance with the animal experiments. This work was supported by grants from the China Medical University and the National Science Council (CMU98-NCTU-06, NSC97-2320-B-039-013-MY3).
References (43)
- et al.
Niuchangchih (Antrodia camphorata) and its potential in treating liver diseases
Journal of Ethnopharmacology
(2009) - et al.
Ethanol extracts of fruiting bodies of Antrodia cinnamomea exhibit anti-migration action in human adenocarcinoma CL1-0 cells through the MAPK and PI3 K/AKT signaling pathways
Phytomedicine
(2012) - et al.
Adenosine acts as an inhibitor of lymphoma cell growth: a major role for the A3 adenosine receptor
European Journal of Cancer
(2000) - et al.
Antrodia camphorata inhibits proliferation of human breast cancer cells in vitro and in vivo
Food and Chemical Toxicology
(2008) - et al.
Berberine inhibits SDF-1-induced AML cells and leukemic stem cells migration via regulation of SDF-1 level in bone marrow stromal cells
Biomedicine and Pharmacotherapy
(2008) - et al.
Thrombopoietin/MPL participates in initiating and maintaining RUNX1-ETO acute myeloid leukemia via PI3 K/AKT signaling
Blood
(2012) - et al.
Expected long-term survival of patients diagnosed with acute myeloblastic leukemia during 2006–2010
Annals of Oncology
(2010) - et al.
MMP-9 in B-cell chronic lymphocytic leukemia is up-regulated by alpha4beta1 integrin or CXCR4 engagement via distinct signaling pathways, localizes to podosomes, and is involved in cell invasion and migration
Blood
(2006) - et al.
Cordycepin inhibits protein synthesis and cell adhesion through effects on signal transduction
Journal of Biological Chemistry
(2010) - et al.
Anti-metastatic activities of Antrodia camphorata against human breast cancer cells mediated through suppression of the MAPK signaling pathway
Food and Chemical Toxicology
(2011)
Cytotoxic triterpenes from Antrodia camphorata and their mode of action in HT-29 human colon cancer cells
Cancer Letters
Cdc2-cyclin E complexes regulate the G1/S phase transition
Nature Cell Biology
Use of arsenic trioxide in haematological malignancies: insight into the clinical development of a novel agent
Current Medical Research and Opinion
Ganoderma lucidum extracts inhibited leukemia WEHI-3 cells in BALB/c mice and promoted an immune response in vivo
Bioscience, Biotechnology, and Biochemistry
Chemical characterization and anti-inflammatory effect of polysaccharides fractionated from submerge-cultured Antrodia camphorata mycelia
Journal of Agricultural and Food Chemistry
Gallic acid downregulates matrix metalloproteinase-2 (MMP-2) and MMP-9 in human leukemia cells with expressed Bcr/Abl
Molecular Nutrition and Food Research
Ethanol extracts of fruiting bodies of Antrodia cinnamomea suppress CL1-5 human lung adenocarcinoma cells migration by inhibiting matrix metalloproteinase-2/9 through ERK, JNK, p38, and PI3 K/Akt signaling pathways
Evidence-based Complementary and Alternative Medicine
Akt- and MAPK-mediated activation and secretion of MMP-9 into stroma in breast cancer cells upon heregulin treatment
Molecular Medicine Reports
p53-independent induction of p21 (WAF1/CIP1), reduction of cyclin B1 and G2/M arrest by the isoflavone genistein in human prostate carcinoma cells
Japanese Journal of Cancer Research
Silymarin and silibinin cause G1 and G2-M cell cycle arrest via distinct circuitries in human prostate cancer PC3 cells: a comparison of flavanone silibinin with flavanolignan mixture silymarin
Oncogene
Matrix metalloproteinase-2 and -9 secreted by leukemic cells increase the permeability of blood-brain barrier by disrupting tight junction proteins
PLoS ONE
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2022, Journal of EthnopharmacologyCitation Excerpt :The compounds in AC extracts mainly include triterpenoids, polysaccharides and steroids (Lu et al., 2013; Zhang et al., 2017). It has been reported that AC has a variety of biological activities, including anti-cancer (Chung et al., 2014; Liu et al., 2013a, 2013b), anti-inflammatory (Huang et al., 2014), anti-oxidation (Liu et al., 2017a; Shih et al., 2017), and hepatoprotective effects (Chen et al., 1995; Liu et al., 2017b). AC barely has toxicity and the products of AC have been registered as health care products in China (Ren et al., 2020).
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