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. 2022 Apr 22:2022:5347718.
doi: 10.1155/2022/5347718. eCollection 2022.

Antileukaemic Cell Proliferation and Cytotoxic Activity of Edible Golden Cordyceps (Cordyceps militaris) Extracts

Singkome Tima  1   2   3 , Tawat Tapingkae  4 , Chaiwat To-Anun  4   5 , Parinn Noireung  4   5 , Phikul Intaparn  4   5 , Wantida Chaiyana  3   6 , Jakkapan Sirithunyalug  3   6 , Pawaret Panyajai  1 , Natsima Viriyaadhammaa  1 , Wariya Nirachonkul  1 , Lapamas Rueankham  1 , Win Lae Aung  1 , Fah Chueahongthong  1 , Sawitree Chiampanichayakul  1   2   3 , Songyot Anuchapreeda  1   2   3
Affiliations

Antileukaemic Cell Proliferation and Cytotoxic Activity of Edible Golden Cordyceps (Cordyceps militaris) Extracts

Singkome Tima et al. Evid Based Complement Alternat Med. .

Abstract

Golden cordyceps (Cordyceps militaris) is a mushroom of the genus Cordyceps. It has been used as a food supplement for both healthy and ill people. In this study, the antileukaemic cell proliferation activities of golden cordyceps extracts were examined and compared with standard cordycepin (CDCP) in EoL-1, U937, and KG-1a cells. Wilms' tumour 1 (WT1) protein was used as a biomarker of leukaemic cell proliferation. The cytotoxicity of the extracts on leukaemic cells was determined using the MTT assay. Their inhibitory effects on WT1 protein expression and cell cycle progression of EoL-1 cells were investigated using Western blotting and flow cytometry, respectively. Induction of KG-1a cell differentiation (using CD11b as a marker) was determined using flow cytometry. The golden cordyceps extracts exhibited cytotoxic effects on leukaemic cells with the highest IC50 value of 16.5 ± 3.9 µg/mL, while there was no effect on normal blood cells. The expression levels of WT1 protein in EoL-1 cells were decreased after treatment with the extracts. Moreover, cell cycle progression and cell proliferation were inhibited. The levels of CD11b increased slightly following the treatment. All these findings confirm the antileukaemic proliferation activity of golden cordyceps.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Figure 1
Figure 1
Golden cordyceps. (a) Culture of golden cordyceps (Cordyceps militaris) in a sterile bottle. (b) Chemical structure of cordycepin (CDCP).
Figure 2
Figure 2
Cytotoxic effects of golden cordyceps extracts on (a) U937, (b) EoL-1, and (c) KG-1a leukaemic cell lines. (d) Doxorubicin (Dox) was used as positive control. Data are presented as the mean ± SD of three independent experiments.
Figure 3
Figure 3
Cytotoxic effects of various golden cordyceps extracts on (a) nonproliferating PBMCs and (b) proliferating PBMCs of three healthy volunteers. Data are presented as the mean ± SD of three independent experiments.
Figure 4
Figure 4
Effects of golden cordyceps extracts on WT1 protein expression in EoL-1 cells. (a) The expression level of WT1 protein (upper band) normalised to GAPDH (lower band). The percentage of WT1 protein expression after treatment with golden cordyceps extracts (crude EtOH, EtOAc, EtOH, and Hex) was compared with that after treatment with CDCP and vehicle control (VC). (b) The total number of viable and dead cells following treatment with golden cordyceps extracts was also evaluated. Data are presented as the mean ± SD of three independent experiments. Asterisks (∗) denote significant differences compared to the VC: ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.
Figure 5
Figure 5
Effect of the ethanol fractional extract of C. militaris (EtOH) on EoL-1 cell cycle progression. Cell cycle histograms were obtained after treatment with various concentrations of (a) EtOH fractional extract and (b) standard cordycepin (CDCP). (c) The proportions of EoL-1 cell population in each phase of the cell cycle were analyzed. The data are presented as the mean ± SD of three independent experiments.
Figure 6
Figure 6
Effects of golden cordyceps extracts and CDCP on KG-1a cell differentiation. (a) Expression levels of CD11b in KG-1a cells after treatment with (red peak) or without (blue peak) golden cordyceps extracts. (b) Relative mean fluorescence intensities (MFI) analyzed by comparing with the vehicle control. (c) The total number of CD11b positive cells measured and compared to that of the negative control group. Data are presented as the mean ± SD of three independent experiments.
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