Recombinant FIP-gat, a Fungal Immunomodulatory Protein from Ganoderma atrum, Induces Growth Inhibition and Cell Death in Breast Cancer Cells

Abstract

FIP-gat, an immunomodulatory protein isolated from Ganoderma atrum, is a new member of the FIP family. Little is known, however, about its expressional properties and antitumor activities. It was availably expressed in Escherichia coli with a total yield of 29.75 mg/L. The migration of recombinant FIP-gat (rFIP-gat) on SDS-PAGE corresponded to the predicted molecular mass, and the band was correctly detected by a specific antibody. To characterize the direct effects of rFIP-gat on MDA-MB-231 breast cancer cells, MDA-MB-231 cells were treated with different concentrations of rFIP-gat in vitro; the results showed that this protein could reduce cell viability dose-dependently with a median inhibitory concentration (IC50) of 9.96 μg/mL and agglutinate the MDA-MB-231 cells at a concentration as low as 5 μg/mL. Furthermore, FIP-gat at a concentration of 10 μg/mL can induce significant growth inhibition and cell death in MDA-MB-231 cells. Notably, FIP-gat treatment triggers significant cell cycle arrest at the G1/S transition and pronounced increase in apoptotic cell population. Molecular assays based on microarray and real-time PCR further revealed the potential mechanisms encompassing growth arrest, apoptosis, and autophagy underlying the phenotypic effects.

Natural Bioactive Compounds from Fungi as Potential Candidates for Protease Inhibitors and Immunomodulators to Apply for Coronaviruses

“…Activating macrophages and cytokines [147,148] Grifolan Grifola frondosa Activating macrophages and lymphokines [149] Lentinan Lentinus edodes T-cell-oriented adjuvant [149] PS-G Ganoderma lucidum Activating macrophages and T lymphocytes [135,136] Schizophyllan Schizophyllum commune Activating T cell, increasing interleukin and TNF-α production [150] Terpenoids…”

Identification and Characterisation of a Novel Protein FIP-sch3 from Stachybotrys chartarum

“…Indirectly, FIPs activate the immune system. Of the direct inhibitory effects on cancer cells, some examples include rLZ-8 promoting apoptosis by G1/S phase arrest in human glioblastoma U-251 MG cells [25] and rFIP-gat significantly inhibiting cell growth, causing cell cycle arrest at the G1/S phase, and inducing apoptosis in MDA-MB-231 breast cancer cells [7]. Previous studies have demonstrated that rFIPs (rFIP-nha from N .…”

“…FIPs regulate immunity by stimulating lymphocyte proliferation, inducing T H 1-specific cytokine synthesis (interleukin-2, interferon-γ, and tumour necrosis factor β), and inhibiting anaphylaxis [1]. The anti-tumour effects that are displayed by FIPs refer to the suppression of tumour cell proliferation, promotion of tumour cell apoptosis, the induction of tumour cell cycle arrest, and the inhibition of tumour cell migration [7,8]. FIPs have very broad potential future applications in medical care, and the anti-tumour effects in particular have attracted extensive attention.…”

“…Homologous cloning involves designing primers using known FIPs as templates and cloning FIPs in the same genus, so it is limited to identifying highly homologous proteins (identity>90%), making it difficult to further characterize FIP proteins. Until now, only five FIPs have been identified using the homologous cloning method: FIP-gmi [14], FIP-gsi from Ganoderma sinense [15], FIP-cru from Chroogomphis rutilus [16], FIP-gat from Ganoderma atrum [7], and LZ-9 [17]. Therefore, researchers have shifted away from the extraction or homologous cloning methods for identifying new FIPs and instead search for sequence similarity in the burgeoning genome databases.…”

Immunomodulatory activity of Ganoderma lucidum immunomodulatory protein via PI3K/Akt and MAPK signaling pathways in macrophage RAW264.7 cells

“…FIP-fve has anti-inflammatory effects on OVA-induced airway inflammation and reduces airway remodeling by suppressing IL-17 (12). In spite of a few studies about anti-tumor (13, 14) and immunomodulatory effects (15–17), the mechanism of these activities remain unclear. The relationship between the activation of these proteins, downstream cytokine expression and physiological function represents an active line of investigation.…”