Mushrooms as potent autophagy modulators in cancer therapy: Current evidence and therapeutic prospects.

Introduction
Mushrooms are valued for their culinary appeal and medicinal properties.1 Being rich in vitamins, minerals, and dietary fiber, they serve as an important component of a healthy diet.2 Moreover, mushrooms have been a core part of traditional Chinese medicine for centuries, due to their health-promoting benefits.3
Ganoderma lucidum (Reishi), Lentinula edodes (Shiitake), and Grifola frondosa (Maitake) are well-known for their immune-boosting, antioxidant, and anti-inflammatory properties.4, 5, 6 Pharmacological research has made tremendous strides recently, resulting in the discovery of mushroom-based medications.7 Two such drugs, PSK (polysaccharide-K)—developed from Trametes versicolor (Turkey tail) and Lentinan from Lentinula edodes—have revealed noteworthy promise in cancer intervention.8,9 Mushrooms are promising agents for cancer treatment owing to their unique bioactive compounds, such as polysaccharides, triterpenoids, and lectins.10, 11, 12 These molecules exhibit anti-cancer activity by modulating signaling pathways involved in cancer cell growth, apoptosis, and invasion, underscoring their potential as novel therapeutic agents.13
Cancer, a multifaceted disease, continues to pose a significant global health concern, causing around 10 million deaths in 2020, as per the World Health Organization.14 Globally, cancer types responsible for the highest mortality rate comprise lung, colorectal, liver, stomach, and breast cancers, followed by prostate, cervical, esophageal, bladder, pancreatic, and head and neck cancers, along with non-Hodgkin lymphoma and leukemia.15,16 Conventional cancer treatments—chemotherapy, radiation, and surgery—often achieve efficacy but are frequently associated with immunosuppression, fatigue, and organ toxicity.17,18 These drawbacks highlight the critical need for alternative therapeutic approaches.
Natural products have gained increasing attention in oncology due to their diverse bioactive constituents and relatively low toxicity.19 Of these, mushrooms present a promising option, serving as a reservoir of distinct bioactive molecules that may target diverse pathways involved in cancer progression.20 Mushrooms exhibit anti-cancer activity through apoptosis induction, immune modulation, and anti-angiogenesis in cancer cells.21, 22, 23, 24 Their potential resides in their capacity to enhance the effectiveness of traditional therapeutic approaches while decreasing their side effects, making them an appealing target for research.24
Autophagy plays a dual role in cancer, acting as a tumor suppressor by removing harmful cellular components and as a tumor promoter by supporting cancer cell survival under stress.25, 26, 27, 28 Targeting autophagy offers a promising therapeutic strategy, either by inducing it to trigger cancer cell death or inhibiting it to enhance treatment sensitivity.29, 30, 31, 32 Mushrooms offer a rich, natural source of autophagy modulators with significant potential to exploit this dual role for therapeutic benefits in cancer.33 They play a significant role in this context, as certain bioactive compounds found in mushrooms modulate autophagy. Mushrooms are notable for their bioactive compounds that regulate autophagy: For instance, triterpenoids induce autophagy-mediated cancer cell death,34 whereas cordycepin inhibits autophagy by suppressing the AMP-activated protein kinase (AMPK)/Mammalian target of rapamycin (mTOR)/uncoordinated-51 (Unc-51) like autophagy activating kinase 1 (ULK1) pathway and lysosomal function, thereby enhancing the efficacy of conventional therapies by sensitizing cancer cells.35
The intrinsic ability of mushroom-derived bioactives to selectively modulate autophagy pathways presents a unique therapeutic opportunity, leveraging the natural safety profile of these compounds to enhance the efficacy and synergy of cancer treatments. This review examines mushrooms as autophagy modulators in cancer therapy, highlighting their bioactive compounds, mechanisms, and potential to enhance treatment efficacy while reducing side effects. It explores new research avenues and therapeutic applications, positioning mushroom-derived compounds as promising agents for safer, more effective cancer treatments.

Brief overview of autophagy and its mechanism
Autophagy, a conserved recycling mechanism in all eukaryotes, has three forms in mammals involving macroautophagy, microautophagy, and chaperone-mediated autophagy, all culminating in lysosomal degradation.36,37 Macroautophagy, the most studied form of autophagy, is a basal process that intensifies under stress conditions like nutrient or energy deprivation. It involves the degradation of cytoplasmic components into metabolites used for energy production or biosynthesis, thereby supporting cell survival.37,38 Under typical growth conditions, macroautophagy helps maintain cells by destroying damaged or unnecessary organelles. Therefore, the primary role of macroautophagy is cytoprotection; however, excessive self-degradation can be harmful. This dynamic process involves autophagosome formation, fusion with lysosomes, and degradation of autophagosomal contents by lysosomal hydrolases.39 The recognition of homologs in higher eukaryotes after the elucidation of autophagy-related genes (ATG) in yeast contributed to our molecular understanding of autophagy. A subset of these ATG proteins, referred to as the “core” molecular machinery, is necessary for the formation of autophagosomes.39,40 The core ATG proteins are categorized into four functional subgroups: (1) the ATG1/ULK kinase complex, (2) two ubiquitin-like conjugation systems involving ATG12 and ATG8/LC3 (microtubule-associated protein 1 light chain 3), (3) the class III phosphatidylinositol 3-kinase (PI3K)/vacuolar protein sorting 34 (Vps34) complex I, and (4) two transmembrane proteins, ATG9/m ATG9 and vacuole membrane protein 1 (VMP1), along with regulatory partners such as ATG18/WD repeat domain phosphoinositide-interacting protein 1 (WIPI1) that facilitate ATG9 trafficking. The ULK complex, consisting of ATG13, ULK1/2, focal adhesion kinase (FAK) family-interacting protein of 200 kDa (FIP200), and ATG101, is essential for the initiation of autophagy.39,41 By determining the level of adenosine monophosphate (AMP) and adenosine triphosphate (ATP), AMPK functions as an energy-sensing kinase that promote autophagy.42,43 Additionally, Ca2+ signaling and endoplasmic reticulum (ER) stress play crucial roles in regulating autophagy by influencing AMPK and Beclin1 activity.44 The Beclin1 complex, which includes Beclin1, Vps34, Vps15, and ATG14, is essential for initiating phagophore formation through generating phosphatidylinositol 3-phosphate (PI3P), a key lipid that recruits proteins necessary for vesicle expansion.45 During nutrient starvation or energy depletion, the AMPK is induced, triggering the ULK1/2 complex.46 This complex further stimulates the Beclin1 complex, inducing PI3P-mediated phagophore, which is nucleated thereafter. Following maturation, autophagosomes fuse with lysosomes to form autolysosomes, where lysosomal hydrolases degrade the cargo. This fusion is regulated by LC3, Ras-related protein Rab-7a (RAB7), pleckstrin homology domain containing family M member 1(PLEKHM1), and the homotypic fusion and vacuole protein sorting (HOPS) complex. The resulting amino acids, fatty acids, and nucleotides are recycled for biosynthesis and energy production.47,48 Moreover, autophagy is induced when activated AMPK phosphorylates the tuberous sclerosis complex (TSC) and reduces the mTOR activity.49,50 As mTOR prevents autophagy, mTOR suppressors have been designed to induce autophagy. Moreover, ATG4 cleaves the precursor proLC3 to form LC3-I, whereas ATG7, ATG3, and the ATG12-ATG5-ATG16 complex conjugate the phosphatidyl-ethanolamine (PE) phospholipid to form LC3-II. The ATG12–ATG5–ATG16L1 complex, the class III phosphatidylinositol 3-kinase (PtdIns3K) complex, LC3–II, and ATG9 are devoted to the phagophore.51, 52, 53 Subsequently, the elongating phagophore engulfs cytoplasmic material to form a double-membraned autophagosome, with LC3-II dissociating from its outer membrane. Fusion with a lysosome creates an autolysosome, where contents are degraded and recycled back into the cytoplasm for reuse [Figure 1].54
In summary, upstream regulators such as AMPK, mTOR, calcium signaling, and ER stress act as metabolic and stress sensors that determine autophagy initiation. These signals act on ULK1 and Beclin1 complexes. AMPK activates ULK1 and inhibits mTOR, lifting its suppression. The Beclin1-Vps34-ATG14 complex produces PI3P, whereas ATG13, FIP200, and VPS15 serve as scaffolds in the cascade.51, 52, 53
Following upstream activation, ATG12-ATG5-ATG16L1 conjugation drives isolation membrane elongation, whereas LC3-II marks autophagosomes for cargo loading and fusion. RAB7, PLEKHM1, and the HOPS complex mediate autophagosome–lysosome fusion, ensuring autophagic flux and degradation by lysosomal hydrolases. These mechanisms highlight potential therapeutic targets in autophagy-related diseases.47,48

Autophagy’s Dilemma: “tumor suppressor or supporter?”
Autophagy, a cellular recycling mechanism, exerts tumor-suppressive or cancer-promoting effects depending on cancer type, stage, and genetic background [Figure 2].55 In early tumorigenesis, autophagy maintains cellular quality by removing damaged organelles and proteins, preserving genomic stability, activating tumor suppressors, regulating PI3K/Akt signaling, and preventing oxidative stress to inhibit cancer development.56 Later, tumor cells exploit autophagy to survive hypoxia, nutrient shortage, and chemotherapy, supporting their growth and metabolic needs.57
Ganoderic acid DM (GA-DM) from G. lucidum illustrates autophagy’s dual role in prostate cancer. It suppresses tumors by upregulating Beclin-1 and ATG5, inducing apoptosis via B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X (Bax) and caspase-3, and boosting ER stress and immune response. However, under prolonged stress, GA-DM can shift autophagy to a survival mechanism, promoting cancer cell protection and highlighting its paradoxical effects in cancer therapy.58

Tumor suppression: role of autophagy in preventing cancer progression
Autophagy suppresses tumor development by reducing cell proliferation, relieving cellular stress, and preserving genomic stability through the clearance of damaged components.59 Key genes like Beclin1 play a vital role; its monoallelic loss leads to spontaneous tumors in mice and is frequently observed in human breast, ovarian, and prostate cancers.60 Beclin1 activates autophagy via Vps34 interaction, reinforcing its tumor-suppressive function.61 Conversely Impaired autophagy is linked to the progression of several cancers, including lung, liver, colorectal, and gastric cancer.62,63
Defective autophagy leads to the accumulation of p62/sequestosome 1 (SQSTM1) aggregates, damaged mitochondria, and misfolded proteins, causing increased reactive oxygen species (ROS) production. This triggers DNA damage and genomic instability, driving tumor initiation and progression. Moreover, impaired autophagy disrupts endoplasmic reticulum (ER) homeostasis, causing persistent ER stress from accumulated unfolded and misfolded proteins.64,65 Chronic unfolded protein response (UPR) activation without functional autophagy intensifies cellular stress and promotes tumorigenesis. Functional autophagy mitigates ER stress by degrading protein aggregates, restoring ER function, and suppressing tumor-promoting signals. Calcium (Ca2+) dynamics are closely tied to autophagy, as the ER is a major Ca2+ reservoir. Impaired autophagy disrupts Ca2+ signaling, causing abnormal cytoplasmic release that activates pro-tumorigenic pathways and elevates oxidative stress, promoting cancer progression. Restoring autophagy rebalances Ca2+ levels, reducing oxidative stress and cellular damage.66 Additionally, reducing p62/SQSTM1 levels in autophagy-deficient cells lowers ROS production and DNA damage. In lung cancer models, p62/SQSTM1-deficient animals resist Ras-induced tumors, unlike wild-type controls. Accumulation of p62/SQSTM1 and increased ROS thus links impaired autophagy to cancer progression.67
Furthermore, other autophagy-related genes, such as ATG5, ATG7, and ATG4C, have vital tumor-suppressive roles in liver, pancreatic, gastric, and colorectal cancers. Mice deficient in ATG5 or ATG7 develop multiple liver tumors from elevated oxidative stress and mitochondrial damage, highlighting the essential roles of these genes in cellular homeostasis and tumor suppression. Although ATG4C contributes to autophagosome formation, its absence has a less pronounced effect on tumorigenesis compared to ATG5 and ATG7, likely due to the presence of compensatory isoforms.68 Additionally, dysregulation of signaling pathways like PI3K/Akt underscores autophagy’s role in tumor suppression. In gliomas and breast, lung, and prostate cancers, PI3K/Akt overactivation suppresses autophagy and promotes tumor growth. PTEN loss or Akt activation reduces autophagy, causing p62 accumulation, increased oxidative stress, and poor prognosis. For instance, in glioma cells, PTEN loss or Akt activation diminishes autophagic activity, thereby exacerbating tumorigenesis by increasing cellular stress and genomic instability.69
Autophagy also plays a role in blocking tumor growth by reducing necrosis and chronic inflammation, which are associated with cancer progression in melanoma, colorectal, pancreatic, and gastric cancers. Autophagy clears damaged cells, preventing the release of pro-inflammatory factors like high mobility group box 1 (HMGB1) that promote a tumor-supportive microenvironment. By reducing necrosis and inflammation, autophagy limits HMGB1 production, thereby inhibiting tumor growth and spread.59, 60, 61

Tumor promotion: how autophagy enables cancer progression
Autophagy acts as a tumor suppressor in early cancer stages but shifts to a pro-survival mechanism in advanced tumors.70 As malignancies grow rapidly and face metabolic stress from poor vascularization and inflammation, autophagy helps cancer cells adapt by supporting survival under nutrient and oxygen deprivation, thereby sustaining tumor growth and its microenvironment.71,72
A key mediator of the inositol-requiring enzyme 1 alpha – X-box binding protein 1 (IRE1α-XBP1) axis promotes autophagy-mediated tumor survival, with IRE1α enhancing tumor cell viability through regulated IRE1-dependent decay (RIDD) of target mRNAs. In this process, IRE1α splices XBP1 mRNA, which induces transcription of pro-survival autophagy genes.65 This also activates tribbles pseudokinase 3 (TRB3), further enhancing autophagy to protect tumor cells from harsh microenvironments, contributing to therapy resistance by enabling cancer cells to evade apoptosis post-chemotherapy and triggers autophagic cell death though excessive activation of ATF4 pathway.73
The protein kinase RNA-like endoplasmic reticulum kinase (PERK)–eukaryotic initiation factor 2 alpha (eIF2α)–activating transcription factor 4 (ATF4) pathway is a key regulator of autophagy-driven tumor promotion. It enhances autophagy under stress by inducing genes that support nutrient recycling and energy balance, helping cancer cells survive and resist apoptosis. In solid tumors, this adaptive response contributes to treatment resistance, though excessive activation can trigger autophagic cell death.74
The ATF6 pathway promotes tumor progression by inducing autophagy-related genes, supporting cancer cell survival through proteostasis maintenance and relief of ER stress.75 Additionally, site-1 protease (S1P) and site-2 protease (S2P)-mediated cleavage of ATF6 in the Golgi apparatus enhances autophagic flux, enabling tumor cells to evade stress-induced apoptosis—an effect particularly critical in aggressive cancers reliant on ER homeostasis for survival.76
Calcium signaling, which plays a tumor-suppressive role under excessive stress, can support tumor survival under moderate conditions.77 Activation of Calcium/Calmodulin-dependent Protein Kinase (CaMK) and c-Jun N-terminal Kinase (JNK), downstream of ER calcium release, enhances autophagy to promote cancer cell adaptation over apoptosis, enabling stress resistance and tumor survival.78
The pro-survival role of autophagy poses challenges in cancer therapy, as tumors exploit it to resist treatment and recover from damage. Inhibitors like chloroquine and hydroxychloroquine enhance therapy by blocking nutrient recycling.79,80 However, autophagy inhibition can trigger necrosis and inflammation, potentially fostering a tumor-supportive environment.

Impact of mushrooms as autophagy modulators in different types of cancer
Recent studies have shown that extracts from various mushrooms [Table 1] and their bioactive compounds [Table 2] influence cancer cells in vitro and in vivo by modulating autophagy. Moreover, comparative studies of the mushrooms with translational potential are depicted in Table 3.

Lung cancer
Lung cancer is the primary cause of cancer-related mortality worldwide, accounting for 18.4% of all such deaths. In the US, an estimated 236,740 new cases and 130,180 deaths from lung cancer were projected for 2022.81 Recent studies have demonstrated that various autophagy-modulating compounds isolated from mushrooms can inhibit lung cancer cell proliferation through multiple signaling pathways [Figure 3].16
GA-DM, a compound from G. lucidum, inhibits the proliferation of A549 and NCI–H460 NSCLC cells by activating autophagy and apoptosis via the PI3K/Akt/mTOR pathway. Autophagy plays a pro-apoptotic role, as its suppression diminishes the apoptotic effects of GA-DM.82 GaH, a natural prostanoid isolated from G. luteomarginatum, blocked the proliferation of A549 and H1299 cells in a time- and concentration-dependent manner. It activated AMPK and upregulated the expression of p53, Beclin-1, and LC3B, thereby inducing autophagy in A549 and H1299 lung cancer cell lines.83 Antrodin C, from T. camphoratus, reduced proliferation and invasion in SPCA-1 cells by activating autophagy through the Akt-mTOR pathway, independent of AMPK. Autophagy inhibition with chloroquine increased apoptosis, suggesting autophagy had a protective role.84 Wang et al. also reported the cytoprotective role of autophagy in A549 lung cancer cells treated with antrodin C.85 Cordycepin (the primary active component of C. militaris) significantly reduced the cell viability of H1792, H1299, H460, H157, and A549 cell lines in a dose-dependent manner, where it activated pro-apoptotic autophagy and caused autophagy-induced disruption of cellular FLICE-like inhibitory protein, long isoform (c-FLIPL).86 Astragurkurol, a terpenoid from Astraeus hygrometricus, inhibits A549 lung cancer cell growth and migration by inducing autophagy (via AVO formation, beclin-1 and Atg7 upregulation, p62 reduction, and PI3K/Akt pathway inactivation). Autophagy inhibition reduced apoptosis, showing its protective role. It also suppressed tumor progression in an ex vivo xenograft model.87 PN50G, a polysaccharide from P. nebrodensis, inhibited A549 cell proliferation and reduced tumor volume and weight in a dose-dependent manner. It induced autophagy by upregulating beclin 1 and promoting LC3-I to LC3-II conversion.88 PFAP, an anti-tumor protein from Pleurotus ferulae lanzi, suppresses mTOR in NSCLC A549 cells, activating autophagy and increasing the expression of P62, LC3 II/I, and related proteins. It also demonstrated tumor growth reduction in a xenograft mouse model in vivo. GMI, a protein from G. microsporum, triggered autophagic cell death in A549 lung cancer cells by enhancing LC3 conversion, reducing p53, and activating a calcium-mediated pathway. It also inhibited tumor growth and regulated autophagy in an A549 xenograft model in vivo.89 GMI inhibited the PI3K/Akt/mTOR pathway, induced autophagy and apoptosis in multidrug-resistant lung cancer cells, and reduced tumor growth in mice xenografts via autophagy and apoptosis, independent of p-glycoprotein overexpression.90,91 Hsin et al. showed that GMI primarily activated autophagy through the PKB pathway in A549 and CaLu-1 lung cancer cells, with mTOR playing a role in GMI-induced autophagy.92 Latcripin-1 (LP1), a novel antitumor mycoprotein from Lentinula edodes, halted the growth and induced autophagy of A549 cells.93

Colorectal cancer
Colorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer-related deaths, with over 1.8 million new cases and 881,000 deaths reported in 2018. Its incidence is projected to rise by 60% by 2030.94 Several autophagy-modulating compounds isolated from various mushrooms have been shown to inhibit the growth of colorectal cancer cells through diverse signaling pathways [Figure 4].95
G. lucidum triterpene extract (GLT) suppressed the progression of human colon cancer cells (HT-29) and inhibited xenograft tumor growth via autophagy induction. This was accomplished by inhibiting the p38 mitogen-activated protein kinase (MAPK) pathway, increasing Beclin-1 and LC3 expression, and reducing p38 MAPK phosphorylation, thereby achieving cancer inhibition through autophagy.96 The fruiting body extract of G. lucidum (GLE) induced autophagy in HCT 116 colorectal cancer cells by modulating key autophagy markers (Beclin-1, LC3B/LC3A, p-mTOR, ATG5, and total mTOR). This reduced cell proliferation in vitro and decreased tumor weight and volume in vivo, contributing to cancer suppression.97
G. lucidum ribonuclease (GLR) suppressed autophagy in HCT 116 and HT-29 cells by increasing p62, upregulating LC3-I, and downregulating LC3-II, inducing apoptosis and inhibiting cancer cell proliferation. This demonstrates that G. lucidum compounds can differentially modulate autophagy.98 Moreover, G. lucidum polysaccharide (GLP) induced autophagy in cancer cells by increasing LC3-II levels and autophagosome formation while disrupted autophagic flux by blocking autophagosome-lysosome fusion. This blockage led to autophagosome accumulation, triggering apoptosis via the MAPK/ERK pathway and inhibiting tumor growth.95 Besides, Antrodia cinnamomea showed potent anticancer effects in the CRC cell line HCT 116. It induced autophagic cell death by upregulating the endoplasmic reticulum stress marker CHOP and its downstream gene TRB3, leading to dephosphorylation of Akt and mTOR and a significant suppression of the tumor growth in vivo in CRC.99
Antrodia salmonea induced autophagy in SW620 colon cancer cells by inhibiting Akt/mTOR signaling, NF-κB, β-catenin expression, and modulating autophagy markers (LC3-II accumulation, p62/SQSTM1 activation, ATG4B inactivation, AVO formation, and Beclin-1/Bcl-2 disruption), enhancing autophagy with a cytoprotective effect.100 Selenium nanoparticles derived from Pleurotus tuber-regium induced autophagy in HCT 116 CRC cells by elevating LC3-II and beclin-1 levels and reducing p62/SQSTM1 expression, leading to cancer cell death and highlighting their potential in anti-CRC therapy.101 Amex7, a combination of extracts from seven medicinal mushrooms (Phellinus linteus, Grifola frondosa, Hericium erinaceum, Lentinula edodes, Sparassis crispa, Trametes versicolor, and Cordyceps militaris), elevated p62 and LC3A/B-II expression in HT-29 cells, promoting autophagy and enhancing its anticancer effect.102 Lentinan, derived from Lentinus edodes, induced autophagy in HT-29 cells by reducing p62 and increasing LC3-II levels, mediated by endoplasmic reticulum stress. This led to autophagic cell death, inhibiting CRC in cell and animal models.103

Gastric cancer
Gastric cancer (GC), a leading epithelial malignancy originating in the stomach, is characterized by its biological complexity and influenced by numerous risk factors.104 In 2020, it accounted for approximately 1.09 million new cases and 0.77 million deaths globally.105 Currently, GC ranks as the third leading cause of cancer-related deaths and the fifth most prevalent cancer worldwide.106 Several studies have shown that mushrooms can inhibit GC by inducing autophagy, serving as a potential mechanism of cancer suppression [Figure 5].
A methanolic extract of G. lucidum fruiting bodies inhibited the growth of human GC AGS cell lines by upregulating the autophagy marker LC3-II and enhancing autophagosome formation, as evidenced by monodansyl cadaverine tagging.27 Moreover, the same methanolic extract further induced autophagy in AGS gastric tumor cells by promoting autophagosome formation, enhancing LC3-II levels, and reducing p62 expression, indicating the autophagy-mediated reduction of cell growth.107 Similarly, a cold methanolic extract (−20 °C) of G. lucidum induced autophagy in AGS cells by increasing autophagosome formation, contributing to cancer inhibition.108 Recombinant Lz-8, derived from G. lucidum, induced autophagic cell death through endoplasmic reticulum stress in human GC cells (SGC-7901). This was evidenced by an increase in LC3 levels, at the protein and mRNA levels, supporting its role in cancer suppression through autophagy.109 Additionally, the ethanolic extract of Lactarius deterrimus (LDE) inhibited the growth of AGS cells while triggering cytoskeleton rearrangements that led to autophagy, further suppressing cancer progression.110 Hispidin, a phenolic compound isolated from Phellinus linteus, induces autophagic and necrotic cell death in SGC-7901 and GES-1 cells via lysosomal membrane permeabilization by inhibiting tubulin polymerization, further promoting cancer cell death.111 Furthermore, N6-(2-hydroxyethyl)-adenosine, a derivative from Cordyceps species, promoted autophagic cell death in SGC-7901 cells by upregulating LC3-II, downregulating p62, and increasing the levels of ATG5, ATG12, and Beclin1, further indicating autophagy-mediated apoptosis and cancer suppression.112 LP1, derived from Lentinula edodes, also triggered autophagy in human GC cells by promoting the formation of autophagosomes and converting LC3I into LC3II, contributing to cancer suppression.113 Additionally, latcripin-7A, another peptide from Lentinula edodes, inhibited the growth of SGC-7901 and BGC-823 cells by suppressing the PI3K/Akt/mTOR pathway, thus inducing autophagy and cancer inhibition.114

Breast cancer
Breast cancer is one of the most prevalent cancers globally, accounting for approximately 25% of all cancer cases in women and contributing to about 15% of cancer-related deaths among women.115,116 In 2023, the United States reported an estimated 300,590 new cases and 43,700 deaths due to breast cancer.117 Autophagy plays a dual role in breast cancer: it supports tumor survival or induces cell death depending on the cellular context.118 Notably, mushrooms with autophagic-modulating properties effectively inhibit breast cancer cell growth [Figure 6].
The ethanolic extract of Chaga mushroom (Inonotus obliquus), widely used in food and medicine, induces autophagy in breast cancer cell lines MDA-MB-231, MDA-MB-468, and MCF7 by inhibiting the mTOR/S6K pathway.119 The extract stimulated AMPK activation, causing mTOR inhibition, a vital controller of cell growth and proliferation. This suppression induced autophagy, promoting the breakdown of cellular materials and potentially reducing cancer cell growth and survival.120
Trametes robiniophila (Huaier) suppresses the mTOR/S6K signaling pathway, triggering autophagy and degrading cellular components. This process reduces tumor growth and promotes autophagic cell death, emphasizing Huaier’s potential as a supportive treatment.121 Furthermore, polysaccharides derived from Trametes robiniophila Murr exhibited tumor-suppressing effects by inducing autophagy. This process led to the degradation of Snail, a critical transcription factor involved in epithelial-mesenchymal transition (EMT). Polysaccharides promoted autophagy, suppressed EMT, and inhibited cancer cell invasion and metastasis, as demonstrated in vivo in Balb/c mice and in vitro in MDA-MB-231 and 4T-1 cancer cells.122
Antrodia salmonea inhibited cancer progression by inducing autophagy and apoptosis. In MDA-MB-231 cells, it increased LC3-II, upregulated ATG7, inhibited mTOR, causing mitochondrial dysfunction, and suppressed tumor growth in xenografted nude mice.123 Moreover, A. salmonea demonstrated its potential to prevent cancer by inducing autophagy, as evidenced in vitro and in vivo. The findings indicated that A. salmonea-induced cytotoxicity involved G2 cell-cycle arrest in MDA-MB-231 cells, characterized by a reduction in cyclin B1, A, and E, as well as CDC2 proteins. The prevention of this cell-cycle arrest by N-acetylcysteine (NAC) suggested that ROS accumulation played a significant role.99 Additionally, A. salmonea treatment decreased COX-2 expression and induced Poly (ADP-ribose) polymerase (PARP) cleavage, which NAC pretreatment reversed, further supporting the involvement of oxidative stress in A. salmonea-mediated cell cycle regulation and tumor suppression in xenografted mice. G. lucidum, dissolved in DMSO, induced autophagy in MCF7 and MDA-MB-231 cells by increasing Beclin-1, LC3, and p62 levels, inhibiting tumor cell proliferation and causing G2 cell cycle arrest, thereby suppressing tumor growth.124 Similarly, combining a small dosage of 5-fluorouracil (5-FU) with ethanol extract from the fruiting bodies of Phellinus linteus, another medicinal mushroom, enhanced the autophagic response in human triple-negative breast cancer (MDA-MB-231) cells. This combination induced cell death through autophagy, enhanced the conversion of LC3-I to LC3-II, promoted the formation of acidic vesicular organelles (AVOs), and facilitated the visualization of numerous double-membraned vacuoles at the ultrastructural level. Combining Phellinus linteus with 5-FU presents a promising strategy for enhancing autophagy-mediated tumor suppression by inhibiting cell proliferation and inducing cell cycle arrest in therapy.125 Additionally, Lentinus edodes showed significant treatment effects through its bioactive polysaccharide Lentinan, which induced autophagy by modulating LC3, p62, and Beclin-1 in tumor tissues of BALB/c-nu mice and MCF7 cells.126 Additionally, latcripin-7A, another bioactive compound from Lentinus edodes, promoted autophagy by increasing the expression of Beclin-1, ATG proteins, and LC3 I/II while decreasing p62 levels in MCF7 and MDA-MB-231 cells.127 Notably, Zhu et al. demonstrated that the bioactive component β-Glucan, derived from the same mushroom, induced autophagic cell death in human breast cancer T47D cells by suppressing Nur77 expression, inhibiting Akt/mTOR signaling, and modulating inflammatory pathways.128 Collectively, Lentinus edodes suppresses breast cancer via autophagy, apoptosis, and inflammation modulation. Similarly, Ganoderma applanatum’s polysaccharides activate MAPK/ERK signaling, inducing autophagy in MCF7 cells, suppressing tumor growth, and demonstrating its therapeutic potential.129

Liver cancer
Liver cancer ranks as the sixth most common cancer worldwide and has a broad geographic distribution, particularly in sub-Saharan Africa, Eastern and Southeast Asia, and Melanesia.130,131 According to GLOBOCAN 2020, primary liver cancer cases are projected to increase by 55%, with a 56.4% rise in related deaths by 2040.132 Mushrooms have been shown to influence liver cancer through the modulation of autophagy, a process that can either suppress tumor development or support cancer cell survival under stress.133
An extract from Hypsizygus marmoreus fruiting bodies induced autophagy-mediated death in Hep 3B liver cancer cells by converting LC3-I to LC3-II and upregulating p62, demonstrating autophagy’s tumor-suppressive role through cellular degradation.134 Moreover, Inonotus baumii extract activates autophagy in liver cancer cells (SMMC-7721) and nude mice via the AMPK/mTOR/ULK1 pathways, increasing LC3-II and decreasing p62, leading to autophagic cell death and tumor growth inhibition.135
Furthermore, G. lucidum spore powder extract exhibited significant anticancer effects in HepG2 and Huh6 cells, as well as in BALB/c mice, by suppressing RACK1 (Receptor for Activated C Kinase 1) O-GlcNAcylation. This suppression led to reduced RACK1 expression and modulated autophagy, malignancy, and immune responses, highlighting the extract’s potential as a therapeutic agent for hepatoblastoma.136 Moreover, cold water extraction from fresh fruiting bodies of Grifola frondosa induced autophagy in 5-week-old male BALB/c athymic nude mice, as well as in human hepatocellular carcinoma (HCC) cell lines Hep3B, HA22T, and Huh7. This effect was mediated through the activation of JNK pathways and the inhibition of PI3K, causing suppressed cell proliferation in vitro and reduced tumor growth in vivo.137,138 Moreover, Grifola frondosa polysaccharide (GEP) combined with vitamin C induced autophagy in SMMC-7721 and HepG2 hepatoma cells by increasing Beclin-1 and LC3II expression and inhibiting the PI3K/Akt/mTOR/p70S6K signaling pathways.139135 Additionally, GFP isolated from the same mushroom induced autophagy in Balb/c mice when combined with vitamin C. Here, autophagy was implicated in cancer suppression by disrupting survival pathways and promoting the degradation of cancer cells.138 Moreover, Agrocybe aegerita lectin induced autophagy in liver cancer by promoting LC3II accumulation, EGFP-LC3 puncta formation, AVO development, and autophagosome creation. This autophagy enhanced apoptosis, suppressing cancer cell viability and contributing to tumor growth reduction.140 Furthermore, armillaridin, present in the edible and medicinal mushroom Armillaria mellea, caused autophagy by promoting LC3 aggregation and the conversion of LC3-I to LC3-II in HepG2 HCC, HA22T, and Huh7 cells, which caused cell death.141 Eburicoic acid from Antrodia cinnamomea fruiting bodies suppressed liver cancer by inducing autophagy. It activated phosphorylation of Beclin-1, JNK, and Bcl-2 pathways, triggering ER stress-mediated cell death and promoting autophagic degradation of damaged cancer cells, reducing proliferation.142

Ovarian cancer
Ovarian cancer (OC) ranks as the seventh most common cancer among women worldwide. It is often asymptomatic in its early stages, making it difficult to detect, and is associated with a high mortality rate, ranking among the deadliest gynecologic cancers alongside uterine and cervical cancers. OC primarily affects women aged 65 and older.143 Globally, it is estimated to cause over 150,000 deaths annually, while in the US, approximately 13,270 deaths were projected in 2023.144,145
Yang et al. demonstrated that bioactive compounds—ergostanes, lanostanes, naphthoquinones, and polyphenols—from the hyphae of Antrodia salmonea aqueous extracts triggered autophagy-mediated cell death in OC cells (SKOV-3 and A2780). This was evidenced by increased LC3-II, GFP-LC3 puncta, and AVO formation, along with p62 activation, ATG4B suppression, ATG7 upregulation, and Beclin-1/Bcl-2 disruption, thereby inhibiting cancer progression and autophagy resistance.146 Similarly, grifolin, a secondary metabolite isolated from the edible mushroom Albatrellus confluens (Northern truffle), stimulates autophagy-mediated tumor suppression in OC cell lines SKOV-3 and A2780 by inhibiting key components of the Akt/mTOR pathway, including p-Akt, p-mTOR, p-p70S6K, and p-4E-BP1, while upregulating autophagic biomarkers LC3B, ATG7, and Beclin-1.147 Ma, Rui et al. found that poricoic acid A from Poria cocos inhibits mTOR/p70S6K signaling, increases LC3-I/II levels, promotes autophagy, and suppresses viability, migration, and invasion of SKOV-3 cells, besides reducing tumor weight in mice.148 However, in human OC cell lines A2780, OVCAR3, SKOV3, TOV112D, and HEK293, cordycepin, an adenosine derivative extracted from C. militaris, can cause autophagic cellular destruction through the ENT1-AMPK-mTOR signaling cascade. Additionally, the methanolic extract of C. militaris, along with cordycepin, demonstrates in vivo efficacy in BALB/c nude mice models by activating AMPK via ENT1 transport.149 This activation leads to downstream autophagic cell death, thereby contributing to the suppression of ovarian cancer. Notably, Coenzyme Q from Antrodia camphorata induces autophagy in SKOV-3 cells as a cytoprotective response, marked by increased LC3-II, GFP-LC3 puncta, AVOs, and Beclin-1/Bcl-2 disruption. This pro-survival autophagy may reduce CoQ’s therapeutic efficacy by helping cancer cells resist apoptosis.150

Cervical cancer
Cervical cancer is the fourth most common cancer in women and the third leading cause of cancer-related deaths in low- and middle-income countries.151 The United States recorded an expected 13,820 new cases and 4360 deaths of cervical cancer by 2024.152
For instance, the aqueous extract of Sanghuangporus baumii inhibits tumor growth in U14 cervical cancer cells in female Kunming mice by upregulating autophagy-related genes (GABARAP, VMP1, VAMP8, and STX17) and increasing LC3II/LC3I ratios.153 The ethanolic extract of Lenzites betulina promotes autophagy in HeLa cells, causing cell cycle arrest, reduced invasion, and tumor growth inhibition; its compound 4′’-hydroxy-6-methoxyaurone reduces drug resistance by targeting P-glycoprotein.154 Additionally, water-soluble extracts from the mycelium of Pleurotus ostreatus and Pleurotus eryngii induce autophagic cell death in SiHa cervical cancer cells via the ER stress–mitochondrial pathway.155 These mushrooms utilize various autophagy mechanisms to suppress cervical cancer progression.

Glioblastoma
In glioblastoma, mushroom compounds like cordycepin, cordycepic acid, and Coenzyme Q10 regulate autophagy differently. Cordycepin from Cordyceps activates autophagy in SH-SY5Y and U-251 cells, promoting cancer cell survival by helping them evade apoptosis through increased LC3I/II expression.156 Conversely, Cordyceps militaris and its mycelial fermentation induce autophagy and apoptosis in GBM8401 and U-87MG glioblastoma cells by downregulating Bcl-2 and inhibiting the Akt-mTOR pathway, using autophagy to suppress cancer and promote cell death.157 Similarly, Coenzyme Q0 from Antrodia camphorata induces autophagy and apoptosis in U87MG and GBM8401 glioblastoma cells by inhibiting the PI3K/Akt/mTOR pathway and triggering ROS-mediated cytotoxicity.158 Thus, although cordycepin initially promotes cancer cell survival by regulating autophagy, Cordyceps militaris and Coenzyme Q10 exploit autophagy to induce apoptosis and suppress glioblastoma, showcasing the dual role of autophagy in cancer progression and therapy.

Skin cancer
Mushroom-derived compounds suppress skin cancer by regulating autophagy. Trametes versicolor extract induces autophagy in SK-MEL-5 melanoma cells, increasing LC3-II and immune checkpoint expression while inhibiting migration.159 GA-DM from G. lucidum promotes autophagy and apoptosis in melanoma cells by boosting Beclin-1 and LC3.160
Cordyceps militaris, delivered via nanoparticles, stimulates autophagy to reduce oxidative stress and support fibroblast regeneration, potentially inhibiting cancer growth.161

Prostate cancer
Cordycepin induces autophagy in LNCaP prostate cancer cells, as evidenced by increased LC3 puncta formation, accumulation of LC3-II, and enhanced autophagic flux. When autophagic flux is blocked by bafilomycin A1, the cells undergo apoptotic cell death, suggesting that cordycepin-induced autophagy functions as a survival mechanism in these cancer cells.162 Resveratrol, a bioactive compound derived from the fruiting bodies of Pleurotus florida, induces autophagy-mediated cell death in prostate cancer cell lines DU145 and PC3. This effect occurs through the depletion of ER calcium stores and inhibition of store-operated calcium entry (SOCE), leading to AMPK activation and suppression of the AKT/mTOR signaling pathway.163 GA-DM, a compound from G. lucidum, suppresses prostate cancer by simultaneously inducing autophagy and apoptosis via Beclin-1, Atg5 upregulation, along with modulation of the Bax/Bcl-2 ratio. It also enhances immune responses through the upregulation of HLA class II molecules. However, under conditions of ER stress, excessive autophagy may paradoxically support tumor survival.164

Leukemia
Ganoderma tsugae ethanolic extracts induce protective autophagy in K562 leukemia cells by increasing LC3-II expression, disrupting the Beclin-1/Bcl-2 complex, and promoting the formation of AVOs. While this suppresses cell viability, it may also contribute to cancer cell survival through autophagy.165 In contrast, poricoic acid A from Poria cocos induces autophagy in T-cell acute lymphoblastic leukemia (T-ALL) cells by modulating the AMPK/mTOR and LC3 signaling pathways, causing reduced cancer cell viability and cell cycle arrest.166 Additionally, a polysaccharide extract from Inonotus taiwanensis inhibits autophagy in THP-1 leukemia cells by decreasing LC3-II levels. This inhibition enhances WSPIS-induced apoptosis and prevents leukemia cells from utilizing autophagy as a survival mechanism.167

Pancreatic cancer
Inhibiting autophagy using mushroom-derived compounds shows promise for treating advanced pancreatic cancer, where cells rely on autophagy for survival and chemotherapy resistance. 4-Acetylantroquinonol B (4-AAQB) from Taiwanofungus camphoratus, Fudan-Yueyang from G. lucidum, and antroquinonol from Antrodia camphorata inhibit autophagy and enhance cancer cell death. 4-AAQB also boosts gemcitabine efficacy by blocking receptor for advanced glycation end-products (RAGE)/HMGB1-mediated PI3K/Akt/MDR1 signaling in MiaPaCa-2 cells.168 Although Fudan-Yueyang prevented autophagosome-lysosome fusion in PANC-1 and BxPC-3 cells, leading to apoptosis.169 Antroquinonol blocked mTOR activity through the PI3K/Akt pathway in PANC-1 and AsPC-1 cells, inducing apoptosis and senescence.170 Although autophagy initially suppresses tumors by maintaining homeostasis, it later contributes to cancer growth and therapy resistance. Targeting autophagy inhibition with mushroom-derived compounds suppresses pancreatic cancer progression and improves conventional treatment efficacy, offering a promising therapy.

Head and neck cancer
In head and neck cancer, Antrodia salmonea suppresses tumor growth by regulating autophagy. In head and neck squamous cell carcinoma (HNSCC) cells with Twist overexpression, A. salmonea fermented broth increases LC3-I/II, acidic AVO formation, and p62 expression, promoting autophagy-induced apoptosis and reducing tumor size in xenografted mice.171 For example, the medicinal mushroom Antrodia cinnamomea contains YMGKI-1, a maleic and succinic acid derivative isolated from its mycelia. YMGKI-1 induces autophagic cell death in head and neck cancer-initiating cells (HN–CICs) by activating AMPK and inhibiting the PI3K-mTOR pathway. This causes a dose-dependent increase in AVOs and an elevated LC3-II/LC3-I ratio, thereby promoting autophagic cell death.172 Additionally, Antrodia camphorata produces Coenzyme Q10, promoting autophagic cell death in FaDu-TWIST1 cells via LC3-II accumulation and AVO formation, reducing tumor growth in mice.173

Urothelial cancer
In urothelial cancer, Ganoderma tsugae’s FIP-gts combined with chloroquine induces autophagy-dependent, caspase-independent cell death, causing autophagosome accumulation and LC3-II activation, resensitizing cisplatin-resistant cancer cells to treatment.174

Esophageal cancer
Finally, in esophageal squamous cell carcinoma (ESCC), ganoderic acid D from G. lucidum induces autophagic cell death in EC9706 and Eca109 cells by downregulating phosphorylated proteins in the PI3K/Akt/mTOR pathway and enhancing autophagosome formation, offering a promising therapeutic approach through synergistic autophagy and apoptosis.175

Safety evaluation of key compounds
Mushroom-derived compounds modulate autophagy in cancer but face clinical challenges like safety and variable efficacy. Cordycepin shows dose-dependent hepatotoxicity, inducing autophagic death in lung cancer but promoting survival in prostate cancer.145,146,148,157,158,160 GA-DM demonstrates selective cytotoxicity but may promote tumor survival under severe endoplasmic reticulum stress, particularly in prostate models.160,147 Antrodin C induces cytotoxic autophagy in colorectal cancer while exhibiting cytoprotective effects in lung adenocarcinoma, which diminishes chemotherapy sensitivity.114,132 Coenzyme Q from Antrodia camphorata shows low toxicity but promotes ovarian cancer survival via autophagy.150,143 Conversely, hispidin shows limited efficacy in TP53-mutant gastric cancers despite inducing lysosomal permeabilization. Lentinan unexpectedly enhances transforming growth factor-β signaling in triple-negative breast cancer, although poricoic acid A promotes tumor growth in immunosuppressed pancreatic cancer models. The identified contradictions, frequently arising from genetic heterogeneity, autophagy flux thresholds, and variations in extract purity, underscore the necessity for standardized dosing, genetic stratification, and combinatorial strategies to enhance therapeutic applications.

Limitations and future perspective
This review underscores the potential of medicinal mushrooms to modulate autophagy as a strategy for cancer prevention, while emphasizing limitations such as insufficient research and dependence on crude extracts, which compromise the clarity and reproducibility of observed anticancer mechanisms. The use of whole mushrooms or unrefined extracts, rather than purified bioactive compounds and standardized formulations, hinders consistency, reproducibility, and the establishment of reliable dose-response relationships.
A major gap exists in comprehensive clinical trial data and large-scale epidemiological studies needed to substantiate the efficacy, optimal dosing, and safety of mushroom-based interventions for cancer prevention and treatment in humans. Additionally, translating promising in vitro and in vivo findings into effective clinical applications remains challenging due to issues such as poor bioavailability, interspecies metabolic differences, and the complexity of human cancer biology compared to model systems. Addressing these limitations is essential for accurately assessing the therapeutic potential of medicinal mushrooms in oncology.
While the modulation of autophagy by mushroom-derived compounds presents a promising avenue for cancer therapy, progress requires coordinated multidisciplinary efforts. Key priorities include standardizing extract preparation, identifying and characterizing active constituents, elucidating mechanisms of action, and conducting rigorous clinical trials to evaluate efficacy and safety. Establishing robust cultivation, extraction, and quality control protocols is critical to ensure the consistency, potency, and safety of mushroom-derived products used in clinical settings.
Advanced researches employing phytochemical and molecular biology techniques are needed to clarify how these compounds influence autophagy and interact with key cancer signaling pathways. Exploring the synergistic potential of combining mushroom-derived autophagy modulators with conventional chemotherapeutics, targeted therapies, synthetic analogs, and immunotherapies may offer a comprehensive and integrative approach to cancer treatment. Such combination strategies could involve administering standardized mushroom extracts as adjunct oral agents alongside conventional treatments, with dosing regimens aligned to therapeutic cycles to enhance efficacy and reduce adverse effects.
Despite encouraging preclinical evidence, the clinical translation of mushroom-derived therapeutics remains in early stages. Well-designed clinical trials are imperative to validate therapeutic efficacy, determine optimal dosing strategies, and assess safety across various cancer types and stages. Personalized treatment approaches, tailored to individual genetic and metabolic profiles, may further enhance therapeutic outcomes and minimize adverse effects.

Conclusion
Medicinal mushrooms have long been recognized for their health benefits and are now undergoing rigorous scientific evaluation for their anticancer properties. Their growing prominence in cancer therapy is largely attributed to a diverse array of bioactive compounds. This review synthesizes evidence from multiple studies demonstrating that various polysaccharides and chemical constituents—such as GA-DM and cordycepin—derived from different mushroom species, induce autophagic cell death, thereby reducing cancer cell viability and proliferation.
Extracts from mushrooms, including Ganoderma lucidum, Antrodia salmonea, and Antrodia cinnamomea, have shown promising effects in tumor suppression, enhanced survival rates, and improved immune responses. Notably, mushroom-derived compounds have exhibited selective cytotoxicity, promoting autophagic cell death in cancer cells while sparing normal cells, highlighting their potential as targeted therapeutic agents.
The development of mushroom-based cancer therapeutics faces several key challenges. These include the standardization of extract preparation, isolation, and characterization of active compounds, and integration with conventional chemotherapeutic regimens. Furthermore, well-designed clinical trials are essential to validate the safety and efficacy of these agents in oncology.
If these challenges are successfully addressed, mushroom-derived therapies may become a valuable component of cancer treatment, offering a natural and effective means to complement existing modalities and improve patient outcomes.

Authors contribution
Md. Mahmudul Hasan: conceptualization, writing–original draft, data extraction, and data analysis, review and editing, ; Eva Azme: conceptualization, writing–original draft, data extraction, and data analysis, review and editing; Rashedul Alam: conceptualization, writing–original draft, data extraction, and data analysis, review and editing; Md. Jahirul Islam Mamun: writing–original draft; Md. Tanvir Chowdhury: writing–original draft; Md. Hossain Rasel: writing–original draft; Md. Safayat Hossen Momen: writing–original draft; Neamul Hoque: writing–original draft; Md. Ekramul Haque Ekram: writing–original draft; Nazmul Hasan Eshaque: writing–original draft; Shakil Ahmed: writing-original draft; Md. Tashrif Rahman Tipu: writing–original draft; Sanjida Shahid Juthi: writing-original draft; Mohammad Fazlul Kabir: writing–original draft; Ahsan Ullah: writing–original draft; Md. Liakot Ali: conceptualization, writing-original draft, manuscript revision. S.M. Moazzem Hossen and Hea-Jong Chung: Conceptualization, supervision, project administration, and manuscript revision. All the authors critically revised and approved the final version of the manuscript.

Ethics statement
None.

Data availability statement
The datasets used in the current study are available from the corresponding author on reasonable request.

Declaration of Generative AI and AI-assisted technologies in the writing process
The authors declare that generative artificial intelligence (AI) and AI assisted technologies were not used in the writing process or any other process during the preparation of this manuscript.

Funding
None.

Conflict of interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.