ArticlePDF AvailableLiterature Review

Characterisation of Extracts and Anti-Cancer Activities of Fomitopsis pinicola

February 2020
Nutrients 12(3):609

DOI:10.3390/nu12030609

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Fomitopsis pinicola (Sw. Karst) is a common bracket fungus, with a woody texture. It is found predominantly in coniferous forests in temperate regions throughout Europe and Asia. Fomitopsis pinicola has been extensively used for medicinal purposes, particularly in Chinese and Korean traditional medicine. In this mini-review, the anti-cancer characteristics of F. pinicola extracts were investigated. In vitro experiments revealed the pro-apoptotic, anti-oxidant and anti-inflammatory properties of extracts, whilst two of three in vivo studies reported an inhibition of tumour growth and prolonged survival. Only studies wherein fungal specimens were sourced from Europe or Asia were included in this review, as samples sourced as F. pinicola from North America were probably not F. pinicola, but a different species. Although not one of the most revered fungal species, F. pinicola has been used as a medicinal fungus for centuries, as well as consumed as a health food supplement. To date, the results from only three in vivo studies, investigating anti-cancer properties, have been published. Further studies, using comprehensively identified specimens, are required to fully elucidate the anti-cancer properties of F. pinicola extracts.

Basidomes of Fomitopsis pinicola in situ. These specimens were identified as F. pinicola by internal spacer region 2 sequencing [13]. (Permission was obtained from NZFocus to utilise this image.) In 2016, Haight et al. reported on an investigation into the suspected F. pinicola complex [14]. Based on samples collected in North America, Europe and Asia and phenotypically identified as F. pinicola, four distinct species were identified, with only F. pinicola found in Europe and Asia. The other three species were found in different regions of North America [1,14]. For this reason, articles based on samples collected outside of Europe and Asia were not included in this review article. F. pinicola is widely available and has been extensively used for medicinal purposes, particularly in Chinese traditional medicine [15]. However, the use of F. pinicola in Central European folk medicine has been largely forgotten [16]. Like many hardwood bracket fungi, it is believed that F. pinicola specimens were traditionally prepared for consumption as a soup/tea or in alcohol [15]. Although not one of the most revered fungal species, F. pinicola (Sw Karst) has been used as a medicinal fungus for centuries for the treatment of headaches, nausea and liver disease [16], as well as in health food supplements [15,16].

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In vitro studies in which the anti-cancer properties of F. pinicola were investigated.

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Nutrients 2020, 12, 609; doi:10.3390/nu12030609 www.mdpi.com/journal/nutrients

Review

Characterisation of Extracts and Anti-Cancer

Activities of Fomitopsis pinicola

Karen S. Bishop *

Discipline of Nutrition and Dietetics/Auckland Cancer Society Research Centre, School of Medical Sciences,

Faculty of Medicine and Health Sciences, University of Auckland, 85 Park Road, Grafton,

Auckland 1023, New Zealand; k.bishop@auckland.ac.nz; Tel.: +64-9-923-4471

Received: 23 January 2020; Accepted: 24 February 2020; Published: 26 February 2020

Abstract: Fomitopsis pinicola (Sw. Karst) is a common bracket fungus, with a woody texture. It is

found predominantly in coniferous forests in temperate regions throughout Europe and Asia.

Fomitopsis pinicola has been extensively used for medicinal purposes, particularly in Chinese and

Korean traditional medicine. In this mini-review, the anti-cancer characteristics of F. pinicola extracts

were investigated. In vitro experiments revealed the pro-apoptotic, anti-oxidant and anti-

inflammatory properties of extracts, whilst two of three in vivo studies reported an inhibition of

tumour growth and prolonged survival. Only studies wherein fungal specimens were sourced from

Europe or Asia were included in this review, as samples sourced as F. pinicola from North America

were probably not F. pinicola, but a different species. Although not one of the most revered fungal

species, F. pinicola has been used as a medicinal fungus for centuries, as well as consumed as a health

food supplement. To date, the results from only three in vivo studies, investigating anti-cancer

properties, have been published. Further studies, using comprehensively identified specimens, are

required to fully elucidate the anti-cancer properties of F. pinicola extracts.

Keywords: anti-cancer properties; extracts; Fomitopsis pinicola; location; medicinal history; sequence

identification; taxonomy

1. Introduction

Fomitopsis pinicola (Sw. Karst), is a common woody fungus found in coniferous forests in

temperate regions throughout Europe and Asia [1], including the Himalayas [2]. Numerous local

names exist for F. pinicola, such as the Japanese name, which is Tsugasaruno-koshikake [3], and the

English name of red-belted bracket fungus [4]. Fomitopsis pinicola is commonly known as a brown-rot

fungus, characterised by bipolar sexual compatibility and the presence of the phenol oxidase,

tyrosinase (with extracellular oxidase not present) [3]. It has been used in Chinese and Korean

traditional folk medicine as an anti-inflammatory agent and for general well-being.

The fruiting body is fan shaped, has a hard, woody texture, can grow up to 40 cm in diameter

(Figure 1), and is often referred to as the red belt conk. The fruiting body has a glossy appearance and

can be red-brown or a lighter colour depending on the age of the specimen. It grows by adding an

additional layer or tube annually. The fungus is saprobic and can also be parasitic, causing heart rot

in living trees, and brown cuboidal rot in dead trees [2]. Decay fungi such as F. pinicola are often

thought to be symbiotic and this could be due to the presence of fungi and nitrogen fixing bacteria at

the same sites on fir trees [5]. In addition, they help circulate forest nutrients through the decay of

dead tree trunks, although the brown rot residues can remain in the soil for extended periods before

breaking down [6,7]. However, F. pinicola and other brown rot species can also contribute

significantly to forestry loses, particularly at sites where the bark has been damaged as might occur

when branches are removed.

Nutrients 2020, 12, 609 2 of 9

Fomitopsis pinicola, like many other fungi, are predominantly identified phenotypically, but

require molecular biology techniques to confirm the identification. Internal spacer region (ITS)2

sequencing is a suitable method that is routinely used for the correct identification of numerous

species, including F. pinicola. Unfortunately, it can appear phenotypically similar to Ganoderma lingzhi

and other species of the genus Fomitopsis, and therefore it is important to confirm the speciation of

the specimen one is working with prior to publication.

A literature review of the anti-cancer properties of F. pinicola was performed using Embase, Web

of Science and Google Scholar. Articles, published in English, where an in vitro and/or in vivo

approach was implemented to investigate the anti-cancer properties of F. pinicola extracts, were

included. Due to extensive fungal species misidentification [8], taxonomy and means of accurate

identification of F. pinicola were also explored. Search terms included “Fomitopsis pinicola”+ “cancer”

+ “in vivo.” Thereafter “anti-inflammatory” was substituted for “cancer”, and an additional article was

returned. In a similar manner “in vitro” was substituted for “in vivo”, and “Fomitopsis pinicola” +

“taxonomy” were also searched. Pearly growing was implemented. This article is not a systematic

review and, together with the implementation of pearly growing, it was decided not to include numbers

and justification for article inclusion and exclusion.

2. The Taxonomy of F. Pinicola

Fungi are poorly, and sometimes incorrectly, described [8]. More recently, sequence-based

classification and identification (SBCI) has been used to detect and classify environmental fungi and

also to confirm or dispute identification or classification of named specimens. The ITS of rRNA genes

can be PCR-amplified and sequenced, and this method is commonly used for SBCI [8]. Further, 16S

rRNA sequences may also be used for this purpose, but it is regarded as less accurate than ITS

sequencing, as the latter is less highly conserved and is therefore more likely to vary from one species

to another [9]. To help avoid misidentifications, Edgar recommends the sequencing of two rather

than one variable region, which could include V3, V4, V5 and ITS, or full-length 16s rRNA or large

subunit rRNA genes [8,9]. With the integration and standardisation of stand-alone databases, and the

incorporation of phylogenetic trees into pipelines used to identify or name specimens, data will be

easier to incorporate into databases and therefore more likely to be deposited, and easier to access,

thus strengthening the accuracy of fungal identification [8].

F. pinicola, an ancient polypore species, is classified according to the Integrated Taxonomic

Information System [10] as follows:

Kingdom: Fungi

Division: Basidiomycota

Class: Agaricomycetes

Order: Polyporales

Family: Fomitopsidaceae

Genus: Fomitopsis

Species: F. pinicola

Fomitopsis pinicola was originally named in 1810 as Boletus pinicola by Swartz and then transferred

to Fomitopsis by Petter Karsten in 1881 [11,12]. Fomitopsis pinicola (Swartz ex Fr.) P. Karst. (1881) was

also named as Polyporus pinicola Fr. [3] before sequencing was used to clearly define the species. More

recently, Binder et al. performed whole-genome sequencing using a shotgun approach, and classified

F. pinicola in the antrodia clade [7].

Nutrients 2020, 12, 609 3 of 9

Figure 1. Basidomes of Fomitopsis pinicola in situ. These specimens were identified as F. pinicola by

internal spacer region 2 sequencing [13]. (Permission was obtained from NZFocus to utilise this image.)

In 2016, Haight et al. reported on an investigation into the suspected F. pinicola complex [14].

Based on samples collected in North America, Europe and Asia and phenotypically identified as F.

pinicola, four distinct species were identified, with only F. pinicola found in Europe and Asia. The

other three species were found in different regions of North America [1,14]. For this reason, articles

based on samples collected outside of Europe and Asia were not included in this review article.

F. pinicola is widely available and has been extensively used for medicinal purposes, particularly

in Chinese traditional medicine [15]. However, the use of F. pinicola in Central European folk

medicine has been largely forgotten [16]. Like many hardwood bracket fungi, it is believed that F.

pinicola specimens were traditionally prepared for consumption as a soup/tea or in alcohol [15].

Although not one of the most revered fungal species, F. pinicola (Sw Karst) has been used as a

medicinal fungus for centuries for the treatment of headaches, nausea and liver disease [16], as well

as in health food supplements [15,16].

3. Active Ingredients

For centuries, medicinal mushrooms have been used by various cultures to enhance health.

Pharmacologic research into medicinal mushrooms, using in vitro, in vivo and clinical studies, has

been used to identify several health benefits and their associated biological pathways [17]. However,

very little research has been carried out on F. pinicola. A variety of extraction methods, whole extracts,

fractions and compounds isolated from the mushroom, have been tested. Many of these are listed in

Table 1. Studies carried out on specimens sourced from North America were not include (e.g., Liu et

al. [18])

Table 1. Extraction method and fraction or compound detected from F. pinicola specimens.

Citation Extraction method Details of method Fraction/Compound/Concentration

Gao et al. 2017,[19]

Methanol

95% ethanol or

methanol for 8 to

10 hours at room

temperature,

evaporated and

washed in water,

evaporated and

lyophilized.

0.210 µM GAE/mg

Hot water

Heated at 100 °C

for 2 to 3 hours; 4

°C overnight;

centrifuged and

lyophilized.

0.185 µM GAE/mg

Ethyl acetate NT. 0.464 µM GAE/mg

Petroleum ether NT. 0.389 µM GAE/mg

Wu et al. 2014, [20] Ethanol

Extracted three

times with 50%

ethanol or water

for 24 h. The

solutions were

Nutrients 2020, 12, 609 4 of 9

filtered, the solvent

removed by

distillation and the

sample was

lyophilised.

Gao et al. 2017, [19] Chloroform/ethanol

The specimens

were dried and

milled. Thereafter,

the powder was

homogenised in

95% ethanol at 45

°C and subjected to

ultrasonic- assisted

extraction. The

extract was

fractionated with

chloroform,

homogenised in

ethanol,

centrifuged and the

supernatant was

filtered.

Ergosterol (105 µg/mg)

Pachymic acid (35.6 µg/mg)

Dehydroeburicoic acid (2.5 µg/mg).

Kao, 2019;

Kao et al. 2018;

Kao et al. 2016, [13,21,22]

Whiskey/Rice wine

(ethanol)

Submerged in

whiskey or rice

wine for six

months, then

freeze dried. The

whiskey extract

was fractionated

1:1:1:1 (water:

methanol: ethanol:

chloroform).

Aqueous fraction

Organic fraction

(56.4% and 43.6% total weight

respectively)

Yoshikawa et al. 2005, [23] Ethanol

Submerged in 70%

ethanol for six

weeks and

separated into

EtOAc and H2O

portions.

Thereafter, the

EtOAc extract was

fractionated using

silica gel column

chromatography,

and some fractions

were further

fractionated using

an HPLC.

Lanostane triterpenes: Fomitopinic

acid A and B

Lanostanoid glycosides: Fomitoside

A-J

Keller et al. 1996, [24] Dichloro-methane

Lanostenoid

derivative.

Seven triterpenes.

3α-(4-car-boxymethyl-3-hydroxy-3-

methylbutanoyloxy)-lanosta

-8,24-dien-21-oic acid,

polyporenic acid C,

3α-acetyloxylanosta-8,24-dien-21-

oic acid,

ergosta-7,22-dien-3/J-ol,

21-hydroxylanosta-8,24-dien- 3-one,

pinicolic acid A,

trametenolic acid B

and pachymic acid21-oic acid

EtOAc—Ethyl acetate; GAE—gallic acid equivalents; HPLC—high-performance liquid

chromatography; NT—not tested.

Although Table 1 includes the compounds that were detected in F. pinicola using different

extraction methods, the anti-cancer activities were not assessed. Various phytochemicals have been

shown to have specific anti-cancer properties, but it is generally accepted that these compounds

Nutrients 2020, 12, 609 5 of 9

probably act synergistically to achieve an anti-cancer effect [25]. Wang et al. identified ergosterol in

a chloroform extract from F. pinicola and observed anti-cancer properties such as a pro-apoptotic and

inhibition of migration effects [26]. Further, in a study published by Yoshikawa et al., fomitopinic

acids and fomitosides inhibited cyclooxygenase (COX) 1 and 2 activity [23]. Although many of the

compounds detected in F. pinicola have not been assessed in isolation, some of the compounds have

been isolated from other species and found to have anti-cancer properties e.g., gallic acid [27]. Based

on the available evidence, it is not possible to determine exactly which compounds exert the strongest

anti-cancer properties, and further research is required.

4. Anti-Cancer Activities

Medicinal properties of mushrooms, based on hearsay, have been recorded for thousands of

years—for example, Ganoderma lucidum (Lingzhi) has been used for general well-being since before

the 5th century by the Chinese [28]; Formes fomentarius has been used as a potent anti-inflammatory

agent by the Greeks (450 BC) [29]; and puffball mushrooms of the genus Calvatia, have been used for

centuries by Native Americans to promote wound healing [29]. More recently, medicinal mushrooms

have been used as an adjuvant to cancer therapy to enhance the effects of treatment and for the

alleviation of side effects from chemo- and radiation therapy (e.g., nausea) [30]. Furthermore,

numerous clinical trials have been conducted to assess the potential anti-cancer properties of both in-

house and commercially prepared medicinal mushrooms [30]. Fewer than ten in vitro studies on

cancer cell lines have been published, but the number of in vivo publications on F. pinicola are even

more limited.

4.1. In Vitro Studies

Numerous cell culture experiments have been used to investigate the anti-cancer properties of

F. pinicola extracts. These studies have been outlined in Table 2.

Table 2. In vitro studies in which the anti-cancer properties of F. pinicola were investigated.

Citation Cell line* Cancer

type

Type of

extract

Cell

viability

1000

µg/ml

(%)

IC50+ Outcomes

Choi et al. 2007, [31]

N/A N/A Not specified N/A N/A

Increased anti-oxidant

activity

HeLa Cervix Water

Ethanol

70.0

25.0

HO-1 Melanoma Water

Ethanol

98.0

40–45

SNU-354 Liver Water

Ethanol

65.0

35–45

SNU-185 Liver Water

Ethanol

60.0

35–45

SK-

Hep3B Liver Water

Ethanol

<82.0

<50.0

Hep3B Liver Water

Ethanol

<82.0

<40.0

PLC/RF/5 Liver Water

Ethanol

95.0

<40.0

Wu et al.2014, [20]

S-180

(mouse) Sarcoma Water

Ethanol

78.9

17.2

Increased CC3, APAF-1

and C-PARP

HepG2 Hepatoma Water

Ethanol

96.6

28.7

Increased CC3; NT; NT

Nutrients 2020, 12, 609 6 of 9

A549 Lung Water

Ethanol

97.0

7.1

Increased CC3; NT; NT

HCT-116 Colon Water

Ethanol

62.5

12.1

Increased CC3; NT; NT

MDA-

MB-231 Breast Water

Ethanol

60.1

34.1

Increased CC3; NT; NT

Gao et al. 2017, [19]

S-180

(mouse) Sarcoma FPKc NT 36.2

Induced late stage

apoptosis/decrease in

MMP/DNA fragmentation

HL-60 Leukemia FPKc NT 41.0 NT

K562 Leukemia FPKc NT 98.9 NT

U937 Leukemia FPKc NT 34.9 NT

SMMC-

7721 Hepatoma FPKc NT 246.2 NT

Eca-109 Esophageal FPKc NT 169.7 NT

Wang et al. 2014, [26]

SW-480 Colon FPKc NT 190.3

Inhibits cell migration and

induce apoptosis.

SW-640 Colon Ergosterol

FPKc

NT 143.3 Induced cell apoptosis

Kao et al. 2018;

Kao et al. 2016

[21,22]

PC3 Prostate WhE NC NT

Upregulation of pro-

apoptotic genes, and

down-regulation of anti-

apoptotic genes.

Significant changes in

gene expression associated

with cell-cycle pathways,

amongst others.

DU145 Prostate WhE NC NT

Significant changes in

gene expression associated

with cell-cycle pathways,

amongst others.

Yoshikawa et al. 2005, [23] N/A N/A

Ethanol

(fomitopinic

acid and

fomitosides)

Anti-inflammatory activity

in response to COX 1 and

*All cell lines are of human origin, unless otherwise stated. + IC50 was measured at 72 h in µg/ml.

Abbreviations: APAF1-apoptotic peptidase activating factor 1; CC3—cleaved caspase 3; COX-

cyclooxygenase; C-PARP—cleaved-poly ADP ribose polymerase; FPKc—F. pinicola chloroform

extract; IC50—half maximal inhibitory concentration; MMP—mitochondrial membrane potential;

N/A-not applicable; NC—not comparable (reported in µl); NT—not tested/not reported; WhE—

whiskey extract.

Hanahan and Weinberg described various hallmarks of cancer [32], which have enabled us to

study the impact of extracts/compounds on these hallmarks (e.g., evasion of programmed cell death)

and their related pathways, rather than on cancer directly. Underlying these hallmarks are

mechanisms such as inflammation, genome instability and the creation of a tumour

microenvironment [32]. Many of the in vitro studies outlined in Table 2 showed an increase in anti-

oxidant activity[31], increase in apoptosis [19,26] or an upregulation of pro-apoptotic genes [21,22],

and anti-inflammatory activity [23]. In addition, PARP, which is involved in DNA repair, genomic

Nutrients 2020, 12, 609 7 of 9

stability and programmed cell death, increased in response to treatment in a sarcoma cell line [20].

Cell cycle dysregulation is another hallmark of cancer [32] and may be a target of the mechanism of

action of FPKc. This reasoning is supported by in vitro evidence showing the inhibition of cell

proliferation; damage to cell membrane in sarcoma but not healthy cells; the triggering of S-phase

cell cycle arrest; a decrease in MMP and release of mitochondrial cytochrome C [19]. Together, these

in vitro studies show that F. pinicola extracts/compounds have anti-cancer activities which warrant

further investigation.

4.2. In Vivo Studies

A small number of in vivo studies have been performed wherein the anti-cancer properties of F.

pinicola were investigated. These studies are listed in Table 3. In two of the studies S-180 sarcoma cells

were used to induce a xenograft [19,20], and in the remaining study, PC3 prostate cancer cells were

used [13]. The extracts (ethanol and chloroform) were both active against the sarcoma xenograft and

inhibited growth, but the powder obtained from an F. pinicola ethanol extract showed no activity

against the prostate cancer xenograft. The discrepancy in the results is thought to be due to the lack

of bioavailability of the ethanol powder extract, as well as treatment at a later stage of disease [13].

In addition to the three studies described in Table 3, Choi et al. also carried out an animal

experiment whereby rats received 0.83 g/kg of mushroom for two weeks following the administration

of ethanol for two weeks [31]. Glutathione, glutathione peroxidase and catalase were all found to be

significantly higher in the intervention versus the control group [31]. Glutathione is an anti-oxidant

and, together with glutathione peroxidase and catalase, protects the cell against oxidative damage,

and thus can exert an anti-tumour effect.

Table 3. In vivo studies in which the anti-cancer properties of F. pinicola were investigated.

Citation Study

design Treatment Type of

extract

Delivery of

extract Outcomes

Wu et al.2014, [20]

Balb/c male

mice with

S180

xenograft

1.5–5 g/kg; 3

and 7 days

prior to

xenograft

Ethanol

extract

Dietary

supplement

Inhibition of tumour growth

(growth inhibitory ratio = 54%

compared to control) and

prolonged survival (40%

survival in the control group,

and 60%–70% survival in the

intervention groups at day 30).

Gao et al. 2017, [19]

ICR mice

with S180

xenograft

200 mg/kg; 7

days prior to

xenograft

FPKc Inoculated

subcutaneously

Inhibition of tumour growth

(inhibition rate = 47.7%

compared to control) and

prolonged survival (control

group–survival ranged from 12

to 15 days, and the intervention

group, survival ranged from 15

to 19 days.

Kao, 2019, [13]

Rag-1 male

mice with

PC3

xenograft

1 g/kg once

the tumour

had reached

200 mm3

Ethanol-

based

extract as a

powder

Oral gavage Dose was tolerated. No

noticeable effect.

Abbreviations: FPKc—chloroform extract of F. pinicola; ICR—institute of cancer research.

In a rat model with diabetes induced by streptozotocin, F. pinicola treatment decreased glucose

levels, restored insulin levels to nearly normal, and pancreatic tissue damage was ameliorated [33].

The alkali extract was more effective than the water extract at reducing the harmful effects of

streptozotocin induced diabetes [33]. Enhanced glucose uptake, and therefore hyperglycaemia, is a

metabolic characteristic of cancer cells, and therefore the link between diabetes and cancers [34] is not

surprising. Although the in vivo study by Lee et al. focused on the impact of F. pinicola extracts on

diabetes, the benefits could be extrapolated to the treatment of cancers. This hyperglycaemic effect,

in the context of cancers, should be investigated further.

Nutrients 2020, 12, 609 8 of 9

4. Limitations

The most obvious limitation of this review is the lack of certainty surrounding the identity of the

specimens used in the studies we discuss. For example, Gao et al. 2017 state that F. pinicola is

traditionally categorized as Reishi [19], yet Reishi is identified as Ganoderma lucidum (Lingzhi) [28]. It

is therefore unclear as to whether Gao et al. studied G. lucidum or F. pinicola and the method of species

identification is not stated.

Another limitation includes the small number of in vivo studies performed. The fact that only

three in vivo studies have been carried out, whereby the anti-cancer properties of F. pinicola were

investigated, indicates the paucity of data and the need to carry out further studies in different cancer

models.

5. Conclusions

In conclusion, further research is required to characterise the anti-cancer activities of F. pinicola

as there is a paucity of data, particularly from in vivo and clinical studies. It would be useful to

identify the bioactive components of F. pinicola and build on the research performed by Wang et al.

and Gao et al. [19,26]. In particular, care must be taken to correctly identify each specimen using

molecular techniques, prior to experimentation. Like many food components, F. pinicola has the

potential to reduce the risk of disease. The advantage of investigating the anti-cancer benefits of F.

pinicola is that the mushroom is not toxic as shown by anecdotal evidence over centuries, as well as

in vivo studies. In addition, it is widely available and is affordable.

Funding: This research received no external funding

Acknowledgments: Proof reading by Renee Alumasa is acknowledged and appreciated.

Conflicts of Interest: The author declares no conflict of interest.

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Chemical composition and antioxidant activity of Fomitopsis pinicola growing on coniferous and deciduous substrates

Article

Full-text available

Sep 2022

Xylotrophic fungi are widespread in all types of forests. The composition of their primary and secondary metabolites including biologically active compounds depends on species and growth substrate; therefore, fungi can become a valuable raw material for biotechnology, pharmaceuticals , and food industry. Fomitopsis pinicola is one of the common species in Russia and Europe, growing predominantly on Pinus sylvestris L., but in mixed forests of the Urals it is also found on Betula pendula Roth. As metabolism of angiosperms and gymnosperms, and their timber properties are different, growth substrate could affect the chemical composition of fungi fruit bodies; therefore our study aimed at metabolite composition of extracts, obtained from F. pinicola collected from birch and pine, and their antioxidant activity. Qualitative analysis revealed alkaloids, phenolics, and anthraquinones. Saponins were found only in the samples obtained from pine. Thin layer chromatography of extracts revealed the same qualitative composition of phenolics, but their amount was higher on birch-4.2 mg g-1 then on pine-3.1 mg•g-1. In ABTS test extracts showed the same antiradical activity. The metabolomics profile obtained by UHPLC-MS totally revealed 116 compounds, and each fungi sample contained more than 70 of them. Thus, the type of substrate influenced on the profile of metabolites and quantitative composition in F. pinicola fruit bodies.

Bioactive extract of Fomitopsis pinicola rich in 11-α- acetoxykhivorin mediates anticancer activity by cytotoxicity, induction of apoptosis, inhibition of tumor growth, angiogenesis and cell cycle progression

Article

Full-text available

Mar 2021

Fomitopsis pinicola (Sw.:Fr.) P. Karst is used in Chinese and Korean traditional medicine The study was aimed to evaluate the anticancer activity of bioactive extract of F. pinicola, mechanism of activity and to identify the bioactive molecules. Ethyl acetate extract (EAE) of F. pinicola was assayed for cytotoxicity, antitumor activity, inhibition of cancerous skin papilloma, antiangiogenesis, inhibition of cell cycle progression and induction of apoptosis. Results showed significant cytotoxicity (IC50 of 100 µg/mL), inhibited tumor growth at a dose of 500 mg/kg, angiogenesis and cell cycle progression at G1 phase. HPTLC analysis of EAE showed 13 peaks indicating the chemical profile. LC-MS analysis revealed 11-α- acetoxykhivorin (RT 9.966, C14H44O12, MW 644.7095) as the major active chemical component. The results concluded that EAE of F. pinicola possessed significant antineoplastic activity. The observed anticancer activity might be assigned to the major chemical components of the extract.

The beneficial effects of Fomitopsis pinicola chloroform extract on a dextran sulfate sodium-induced ulcerative colitis mice model

Article

Full-text available

Nov 2022

Background: As an intestinal non-specific inflammatory lesion, ulcerative colitis (UC) affects the health of many individuals. This study examined the possible beneficial effects of the chloroform extract of Fomitopsis pinicola (Swartz.: Fr) Karst (FPKc) on UC. Methods: The mice were given free access to drink with 4% dextran sulfate sodium (DSS) for 1 week to establish acute UC model. Next, 35 mg of FPKc or sulfasalazine (SASP) was given to the mice via gavage for 3 weeks. The disease activity index (DAI) and colonic mucosa damage index (CMDI) scores were calculated. The colon tissues of the mice were collected to measure the length and perform hematoxylin and eosin staining. The thymus and spleen indexes were determined. Interleukin (IL)-6, IL-8, tumor necrosis factor-α, aminotransferase (AST) and alanine aminotransferase (ALT) levels in the serum were determined. Results: FPKc or SASP treatment alleviated hematochezia and weight loss, ameliorated DAI and CMDI scores, and improved the crypt structure and length of the colon tissues. Relative to the UC model group, the spleen index in the FPKc group was reduced, which was accompanied by decreases of the IL-6 and IL-8 levels in the serum. FPKc also lowered the AST and ALT levels in the serum of the UC mice. Conclusions: FPKc protected the mice from DSS-induced UC injury. It may be that FPKc activates immune regulation and downregulates the expression of pro-inflammatory cytokines.

Wood-inhabiting macrofungi Hymenochaetales and Polyporales (Basidiomycota) in the Amazon Forest: relationship the abiotic factors and substrate colonization

Article

Full-text available

Oct 2022
AN ACAD BRAS CIENC

Hymenochaetales and Polyporales are important macrofungi for the maintenance of tropical forests, since they act directly in the nutrient cycling of the wood decomposition. In the Amazon, the largest tropical forest in the world, knowledge about Agaricomycetes is still insipient, since many areas have not yet been inventoried and new records appear each new study. To increase ecological knowledge about the Hymenochaetales and Polyporales, in the Brazilian Amazon region, collections were conducted in western Pará, Brazil, relating these fungi to the substrate they colonize and to environmental variables. 91 species were identified, with greater macrofungi richness associated with the rainy season; these fungi showed preferences for dead woods, of small diameter (class 1 = 5,9 ˫ 39 cm) and, in stages of decomposition still rigid or intermediate. The abundance and richness of Hymenochaetales and Polyporales were influenced by air humidity and the assemblage composition was influenced by temperature, air humidity and rainfall. The results indicate a rich diversity for western Pará region, these species are associated with environmental conditions, and may be threatened by the increasing pressure of human activity in the Brazilian Amazon.

Review on health status, chemical composition and antimicrobial properties of the four species of the genus Cedrus

Article

Full-text available

Sep 2022

This article presents a literature review on the chemical composition, antimicrobial activities of cedar oils, and the main diseases this tree is subjected to. The studies included in this review have drawn upon several databases including Scopus, Springer, ScienceDirect, Web of sciences. The keywords used in data collection were C. atlantica M, C. libani L, C. deodara L, C. brevifolia H, Red ring rot (M’jej), Cubic brown rot (Saboune). Genus Cedrus is rich in bioactive molecules such as himachalenes, atlantones and generally terpenes. These molecules have an important medicinal and cosmetic properties maintained by the inhibition and destruction of many bacteria and fungi, along with other several biological activities. Cedar suffers from pests and fungal attacks, which cause two types of fungal decay: the first one is brown cubic rot caused by Fomitopsis pinicola or Ungulina officinalis, and the red ring rot caused by Trametes pini or Phellinus chrysoloma.

Antioxidative and Cytoprotective Effects of Ganoderma applanatum and Fomitopsis pinicola in PC12 Adrenal Phaeochromocytoma Cells

Article

Aug 2022
INT J MED MUSHROOMS

Considering the impact of oxidative stress on the development of many diseases, together with the role of natural antioxidants in maintaining physiological balance in humans, medicinal mushrooms are potential sources of bioactive compounds against many diseases. In the present work, in vitro evaluation of the biological activities of the alcoholic extracts of two wild tree mushrooms, namely, Ganoderma applanatum and Fomitopsis pinicola, has been performed. Extraction of G. applanatum (GAE) and F. pinicola (FPE) was conducted with 60% ethanol and 100% ethanol sequentially. UPLC-MS/MS identification was conducted on the two mushrooms extracts. A total of 15 substances were identified in GAE, including 3 spiro meroterpenoids and 12 triterpenoids; a total of 14 chemical constituents were iden¬tified in FPE, including 8 triterpenoids, 4 triterpene glycosides, 1 lanosterol, and 1 lanostanoid. The resulting extracts were examined for their in vitro antioxidative and cytoprotective effects against AAPH-induced oxidative damage. Our results demonstrated that both extracts have potent antioxidative activities, when GAE was 0.2 mg/mL, the clearance rates of DPPH and ABTS have reached 93.34% and 99.93%, respectively. When FPE was 1.4 mg/mL and 0.6 mg/mL, the scavenging rates of DPPH and ABTS have reached 91.76% and 100%, respectively. Both the alcoholic extracts of G. applanatum and F. pinicola were able to protect the AAPH-induced damage and could effectively inhibit cell aging via β-galactosidase (SA β-gal) staining activity test and scanning electron microscopy analysis.

MEDICINAL MUSHROOM BIOACTIVES: POTENTIAL SOURCES FOR ANTI-CANCER DRUG DEVELOPMENT

Article

Full-text available

Oct 2020

Mushrooms represent a major yet largely untapped source of therapeutically useful bioactive compounds. Despite mushrooms were in use since antiquity in traditional folk medicine attempts to isolate their bioactive components and to elucidate their medicinal properties have started only recently. Many pharmaceutical substances with unique properties were recently extracted from mushrooms and made their way all over the world. A number of medicinal mushrooms have been identified to possess anticancer effects recently. Some of the well-known examples are Lentinan from Lentinus edodes, Krestin from Trametes versicolor, Ganopoly from Ganoderma lucidum and Schizophyllan from Schizophyllum commune. We investigated the anticancer activities of a number of medicinal mushrooms in our laboratory. Some of the recent scientific evdences on the anticancer activities of Ganoderma lucidum, Phellinus rimosus, and Fomitopsis pinicola are discussed in this short review.

Anti-Oxidant and Anticancerous Effect of Fomitopsis officinalis (Vill. ex Fr. Bond. et Sing) Mushroom on Hepatocellular Carcinoma Cells In Vitro through NF-kB Pathway

Article

Jun 2021
Anti Canc Agents Med Chem

Background Fomitopsis officinalis (Vill. ex Fr. Bond. et Sing) is a medicinal mushroom, commonly called ‘Agarikon’, traditionally used to treat cough and asthma in the Mongolian population. Objective The objective of this study was to examine the significance of biological activity of F. officinalis, and evaluate the antioxidant and anticancer activity of six fractions of F. officinalis residues (Fo1-powder form dissolved in ethanol, Fo2-petroleum ether residue, Fo3-chloroformic, Fo4-ethylacetate, Fo5-buthanolic, and Fo6-water-ethanolic) against hepatocellular carcinoma cells. Methods We performed in vitro studies of cell proliferation and viability assay, annexin V-FITC/Propidium Iodide assay, and NF-kB signaling pathway by immunoblot analysis. Results Our findings revealed that all six fractions/extracts have antioxidant activity, and somehow, they exert anticancerous effects against cancer cells. In cancerous cell lines (HepG2 and LO2), Fo3 chloroformic extract promoted the cancer cell apoptosis, cell viability, activated G2/M-phase cell cycle, and selectively induced NF-kB proteins, revealing itself as a novel antitumor extract. Conclusion This study reports that Fo3-chloroformic extract is rich in antitumor activity; it was previously not investigated in cancer. To study the impact of F. officinalis among natural products to treat/prevent oxidative stress disorders or cancers, further examinations are needed. However, this study assessed only one extract, Fo3-chloroformic, which has a significant impact on cancer cell lines.

Antiinflammatory Triterpenoids from the Fruiting Bodies of Fomitopsis pinicola

Article

Dec 2020

Twelve undescribed lanostane-type triterpenes, and twenty-two known triterpenes were isolated and identified from a medicinal bracket fungus Fomitopsis pinicola (Sw.) P. Karst. The structures of these compounds were determined by spectroscopic and spectrometric analyses. The antiinflammatory potential of thirty-two triterpene compounds was evaluated using neutrophils as an assay model, and pinicolasin J was the most potent inhibitor of superoxide anion generation and elastase release, with IC50 values of 1.81 ± 0.44 and 2.50 ± 0.64 μM, respectively. This study provides scientific insight into the nutritional supplement value and medicinal development of Fomitopsis pinicola.

Fomitopsis mounceae and F. schrenkii-two new species from North America in the F. pinicola complex

Article

Full-text available

Mar 2019
MYCOLOGIA

Two new species, Fomitopsis mounceae and F. schrenkii (Polyporales, Basidiomycota) in the F. pinicola species complex in North America, are described and illustrated. Previous molecular phylogenetic analyses identified three well-delimited lineages that represent F. mounceae and F. ochracea from Canada, the Appalachian Mountains, and the northern United States and F. schrenkii from western and southwestern regions of the United States. Fomitopsis pinicola sensu stricto is restricted to Eurasia and does not occur in North America. Morphological descriptions of basidiocarps and cultures for F. mounceae, F. schrenkii, and F. ochracea are presented. The three species are readily differentiated by nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) sequence, geographic distribution, and basidiospore size. Polyporus ponderosus H. Schrenk is an earlier illegitimate synonym of F. schrenkii. Both F. mounceae and F. schrenkii have a heterothallic multiallelic incompatibility system.

Accuracy of taxonomy prediction for 16S rRNA and fungal ITS sequences

Article

Full-text available

Apr 2018

Robert C. Edgar

Prediction of taxonomy for marker gene sequences such as 16S ribosomal RNA (rRNA) is a fundamental task in microbiology. Most experimentally observed sequences are diverged from reference sequences of authoritatively named organisms, creating a challenge for prediction methods. I assessed the accuracy of several algorithms using cross-validation by identity, a new benchmark strategy which explicitly models the variation in distances between query sequences and the closest entry in a reference database. When the accuracy of genus predictions was averaged over a representative range of identities with the reference database (100%, 99%, 97%, 95% and 90%), all tested methods had ≤50% accuracy on the currently-popular V4 region of 16S rRNA. Accuracy was found to fall rapidly with identity; for example, better methods were found to have V4 genus prediction accuracy of ∼100% at 100% identity but ∼50% at 97% identity. The relationship between identity and taxonomy was quantified as the probability that a rank is the lowest shared by a pair of sequences with a given pair-wise identity. With the V4 region, 95% identity was found to be a twilight zone where taxonomy is highly ambiguous because the probabilities that the lowest shared rank between pairs of sequences is genus, family, order or class are approximately equal.

In Vitro and In Vivo Activity of Fomitopsis Pinicola (Sw. Ex Fr.) Karst Chloroform (Fpkc) Extract Against S180 Tumor Cells

Article

Full-text available

Dec 2017

Background/aims: Non-toxic fomitopsis is has been traditionally used in folk medicine in many countries for its anti-inflammatory and anti-vascular disease activities. The present study investigates the antitumor effect of Fomitopsis pinicola (Sw. Ex Fr.) Karst chloroform extract (FPKc) on S180 tumor cells in vitro and in vivo and we determined the underlying mechanisms. Methods: HPLC was employed to analyze the constituents of FPKc. In-vitro 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to quantify the growth inhibition of FPKc; Propidium iodide (PI) exclusion assay and scanning electron microscopy (SEM) were used to observe the damage on the cell membrane and the changes of the cell morphology; Staining with Hoechst 33342/propidium iodide (HO/PI) and the application of the Annexin V-FITC/PI analysis permitted to observe the cell death triggered by FPKc; DNA damage and cell cycle arrest were detected by flow cytometry; Rhodamine 123 (RH123) and Cytochrome C were used as dyes to investigate the alterations of the mitochondria. In-vivo tumor inhibition and mice survival time were determined. Results: The results of the HPLC assay indicated that FPKc might contain DA (dehydroeburiconic acid), PA (pachymic acid), and ES (ergosterol), at percentages of 0.25%, 17.8%, and 10.5%, respectively. Concerning the study of the biological function, the results showed that FPKc exhibited preferential and significant suppression of proliferation on specific cell lines including S180, HL-60, U937, K562, SMMC-7721, and Eca-109 cells, which induced plasma membrane and cell morphology damages, triggering S180 tumor-cells late apoptosis and leading to DNA damage and S phase arrest. Mitochondria were suspected to play a vital role in these changes. In vivo data indicated that FPKc inhibited the solid tumor growth and prolonged the survival time of tumor-bearing mice. Moreover, FPKc provoked only little damage on normal cells in vitro and also on normal tissues in vivo. Conclusion: FPKc inhibited S180 tumor cells growth and prolonged the lifespan of mice. In vitro, we found that FPKc induced S180 tumor cells apoptosis and cell cycle arrest, possibly via the mitochondrial pathway.

Sequence-based classification and identification of Fungi

Article

Full-text available

Oct 2016

Fungal taxonomy and ecology have been revolutionized by the application of molecular methods and both have increasing connections to genomics and functional biology. However, data streams from traditional specimen- and culture-based systematics are not yet fully integrated with those from metagenomic and metatranscriptomic studies, which limits understanding of the taxonomic diversity and metabolic properties of fungal communities. This article reviews current resources, needs, and opportunities for sequence-based classification and identification (SBCI) in fungi as well as related efforts in prokaryotes. To realize the full potential of fungal SBCI it will be necessary to make advances in multiple areas. Improvements in sequencing methods, including long-read and single-cell technologies, will empower fungal molecular ecologists to look beyond ITS and current shotgun metagenomics approaches. Data quality and accessibility will be enhanced by attention to data and metadata standards and rigorous enforcement of policies for deposition of data and workflows. Taxonomic communities will need to develop best practices for molecular characterization in their focal clades, while also contributing to globally useful datasets including ITS. Changes to nomenclatural rules are needed to enable valid publication of sequence-based taxon descriptions. Finally, cultural shifts are necessary to promote adoption of SBCI and to accord professional credit to individuals who contribute to community resources.

Phylogeny of Fomitopsis pinicola: A species complex

Article

Full-text available

Sep 2016

Fungal species with a broad distribution may exhibit considerable genetic variation over their geographic ranges. Variation may develop among populations based on geographic isolation, lack of migration, and genetic drift, though this genetic variation may not always be evident when examining phenotypic characters. Fomitopsis pinicola is an abundant saprotrophic fungus found on decaying logs throughout temperate regions of the Northern Hemisphere. Phylogenetic studies have addressed the relationship of F. pinicola to other wood-rotting fungi, but pan-continental variation within F. pinicola has not been addressed using molecular data. While forms found growing on hardwood and softwood hosts exhibit variation in habit and appearance, it is unknown if these forms are genetically distinct. In this study, we generated DNA sequences of the nuc rDNA internal transcribed spacers (ITS), the TEF1 gene encoding translation elongation factor 1-α, and the RPB2 gene encoding the second largest subunit of RNA polymerase II for collections across all major geographic regions where this fungus occurs, with a primary focus on North America. We used Bayesian and maximum likelihood analyses and evaluated the gene trees within the species tree using coalescent methods to elucidate evolutionarily independent lineages. We find that F. pinicola sensu lato encompasses four well-supported, congruent clades: a European clade, southwestern US clade, and two sympatric northern North American clades. Each clade represents distinct species according to phylogenetic and population-genetic species concepts. Morphological data currently available for F. pinicola do not delimit these species, and three of the species are not specific to either hardwood or softwood trees. Originally described from Europe, F. pinicola appears to be restricted to Eurasia. Based on DNA data obtained from an isotype, one well-defined and widespread clade found only in North America represents the recently described Fomitopsis ochracea. The remaining two North American clades represent previously undescribed species. © 2016 by The Mycological Society of America, Lawrence, KS 66044-8897.

Identification of Potential Anticancer Activities of Novel Ganoderma lucidum Extracts Using Gene Expression and Pathway Network Analysis

Article

Full-text available

Feb 2016

Ganoderma lucidum (lingzhi) has been used for the general promotion of health in Asia for many centuries. The common method of consumption is to boil lingzhi in water and then drink the liquid. In this study, we examined the potential anticancer activities of G. lucidum submerged in two commonly consumed forms of alcohol in East Asia: malt whiskey and rice wine. The anticancer effect of G. lucidum, using whiskey and rice wine-based extraction methods, has not been previously reported. The growth inhibition of G. lucidum whiskey and rice wine extracts on the prostate cancer cell lines, PC3 and DU145, was determined. Using Affymetrix gene expression assays, several biologically active pathways associated with the anticancer activities of G. lucidum extracts were identified. Using gene expression analysis (real-time polymerase chain reaction [RT-PCR]) and protein analysis (Western blotting), we confirmed the expression of key genes and their associated proteins that were initially identified with Affymetrix gene expression analysis.

Fungal strain matters: colony growth and bioactivity of the European medicinal polypores Fomes fomentarius, Fomitopsis pinicola and Piptoporus betulinus

Article

Full-text available

Jan 2015

Polypores have been applied in traditional Chinese medicine up to the present day, and are becoming more and more popular worldwide. They show a wide range of bioactivities including anti-cancer, anti-inflammatory, antiviral and immuno-enhancing effects. Their secondary metabolites have been the focus of many studies, but the importance of fungal strain for bioactivity and metabolite production has not been investigated so far for these Basidiomycetes. Therefore, we screened several strains from three medicinal polypore species from traditional European medicine: Fomes fomentarius, Fomitopsis pinicola and Piptoporus betulinus. A total of 22 strains were compared concerning their growth rates, optimum growth temperatures, as well as antimicrobial and antifungal properties of ethanolic fruit body extracts. The morphological identification of strains was confirmed based on rDNA ITS phylogenetic analyses. Our results showed that species delimitation is critical due to the presence of several distinct lineages, e.g. within the Fomes fomentarius species complex. Fungal strains within one lineage showed distinct differences in optimum growth temperatures, in secondary metabolite production, and accordingly, in their bioactivities. In general, F. pinicola and P. betulinus extracts exerted distinct antibiotic activities against Bacillus subtilis and Staphylococcus aureus at minimum inhibitory concentrations (MIC) ranging from 31-125 μg mL−1; The antifungal activities of all three polypores against Aspergillus flavus, A. fumigatus, Absidia orchidis and Candida krusei were often strain-specific, ranging from 125-1000 μg mL−1. Our results highlight that a reliable species identification, followed by an extensive screening for a ‘best strain’ is an essential prerequisite for the proper identification of bioactive material.

Investigating Migration Inhibition and Apoptotic Effects of Fomitopsis pinicola Chloroform Extract on Human Colorectal Cancer SW-480 Cells

Article

Full-text available

Jul 2014
PLOS ONE

Background Fomitopsis pinicola (Sw. Ex Fr.m) Karst (FPK) which belongs to the Basidiomycota fungal class is one of the most popular medical fungi in China. It has been used for many diseases: cancer, heart diseases, diabetes and so on. However, little study on the pro-apoptotic effect and migration inhibition of FPK chloroform extract (FPKc) has been reported and the possible involved mechanism has not been illuminated. Methodology/Principal Findings Chemical analysis was performed by HPLC which showed ergosterol (ES) concentration was 105 µg/mg. MTT assay revealed that FPKc could selectively inhibit SW-480 cells viability with the IC50 of 190.28 µg/ml. Wound healing and transwell assay indicated that FPKc could inhibit the migration of SW-480 cells obviously, FPKc could also dramatically decreased the matrix metalloproteinases-2, 9 (MMP-2 and MMP-9) expression. Annexin V–FITC/PI staining, nuclear Hoechst 33342 staining and DNA fragmentation analysis revealed that FPKc and ES could induce SW-480 cells apoptosis. The apoptosis process closely involved in ROS accumulation and depletion of GSH, activation of caspase 3, poly (ADP-ribose) polymerase (PARP) degradation. FPKc could also up-regulate P53 expression and thus lead to G1 phase arrest. When SW-480 cells were pretreated with N-acetylcysteine (NAC), the ROS generation, cell viability and apoptotic ratio were partially declined, which indicated that ROS was vertical in the pro-apoptosis process induced by FPKc. Moreover, in the whole process, ES which has been previously found in FPKc had the similar effect to FPKc. Thus we could conclude that ES, as one of the highest abundant components in FPKc, might also be one of the active constituents. Conclusion/Significance FPKc could inhibit the migration of SW-480 cells, induce SW-480 cells G1 phase arrest and cause ROS-mediated apoptosis effect. And ES might be one of the effective constituents in the whole process.

Medicinal Mushrooms: Ancient Remedies Meet Modern Science

Article