title: GMI, an immunomodulatory protein from Ganoderma microsporum, induces autophagy in non-small cell lung cancer cells.
abstract: Autophagy is a self-digestive process that degrades the cytoplasmic constituents. Immunomodulatory protein, one major bioactive component of Ganoderma, has antitumor activity. In this study, recombinant fungal immunomodulatory protein, GMI, was cloned from Ganoderma microsporum and purified. We demonstrated that GMI induces lung cancer cell death by activating autophagy, but does not induce apoptotic cell death. On western blot, GMI increased LC3 conversion and decreased p53 expression in a time- and concentration-dependent manner. Cytoplasmic calcium chelator BAPTA-AM was used to prove that GMI promotes autophagy via a calcium-mediated signaling pathway. 3-methyladenine (3-MA), an autophagy inhibitor, enhanced the cytotoxicity of GMI on cell viability assay. Using VZV-G pseudotyped lentivirus-shRNA system for autophagy-related genes silencing, the capabilities of GMI to reduce cell viability and colony formation were abolished in autophagy-defective cells. Furthermore, GMI did not stimulate apoptosis after blocking of autophagy by 3-MA or shRNA knockdown system. In xenograft studies, oral administration of GMI inhibited the tumor growth and induced autophagy significantly in nude mice that had received a subcutaneous injection of A549 cells. This is the first study to reveal the novel function of GMI in activating autophagy. GMI may be a potential chemopreventive agent against non-small cell lung cancer.
authors.affiliation: Institute of Medical and Molecular Toxicology, Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan ROC.
authors.forename: I-Lun
authors.initials: IL
authors.lastname: Hsin
authors.forename: Chu-Chyn
authors.initials: CC
authors.lastname: Ou
authors.forename: Tzu-Chin
authors.initials: TC
authors.lastname: Wu
authors.forename: Ming-Shiou
authors.initials: MS
authors.lastname: Jan
authors.forename: Ming-Fang
authors.initials: MF
authors.lastname: Wu
authors.forename: Ling-Yen
authors.initials: LY
authors.lastname: Chiu
authors.forename: Ko-Huang
authors.initials: KH
authors.lastname: Lue
authors.forename: Jiunn-Liang
authors.initials: JL
authors.lastname: Ko
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D002289:Carcinoma, Non-Small-Cell Lung / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005656:Fungal Proteins / Q000494:pharmacology*; D032902:Ganoderma / Q000737:chemistry*; D020868:Gene Silencing / Q000187:drug effects; D007155:Immunologic Factors / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000473:pathology*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014410:Tumor Stem Cell Assay; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D005656:Fungal Proteins; D007155:Immunologic Factors; C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins; D016159:Tumor Suppressor Protein p53; C025946:3-methyladenine; D000225:Adenine; D002118:Calcium