title: Secondary Metabolites from Endophytic Fungus Penicillium pinophilum Induce ROS-Mediated Apoptosis through Mitochondrial Pathway in Pancreatic Cancer Cells.
abstract: The endophytic fungus strain MRCJ-326, isolated from Allium schoenoprasum, which is also known as Snow Mountain Garlic or Kashmiri garlic, was identified as Penicillium pinophilum on the basis of morphological characteristics and internal transcribed spacer region nucleotide sequence analysis. The endophytic fungus extract was subjected to 2D-SEPBOX bioactivity-guided fractionation and purification. The anthraquinone class of the bioactive secondary metabolites were isolated and characterized as oxyskyrin (1), skyrin (2), dicatenarin (3), and 1,6,8-trihydroxy-3-hydroxy methylanthraquinone (4) by spectral analysis. Dicatenarin and skyrin showed marked growth inhibition against the NCI60/ATCC panel of human cancer cell lines with least IC50 values of 12 µg/mL and 27 µg/mL, respectively, against the human pancreatic cancer (MIA PaCa-2) cell line. The phenolic hydroxyl group in anthraquinones plays a crucial role in the oxidative process and bioactivity. Mechanistically, these compounds, i.e., dicatenarin and skyrin, significantly induce apoptosis and transmit the apoptotic signal via intracellular reactive oxygen species generation, thereby inducing a change in the mitochondrial transmembrane potential and induction of the mitochondrial-mediated apoptotic pathway. Our data indicated that dicatenarin and skyrin induce reactive oxygen species-mediated mitochondrial permeability transition and resulted in an increased induction of caspase-3 apoptotic proteins in human pancreatic cancer (MIA PaCa-2) cells. Dicatenarin showed a more pronounced cytotoxic/proapopotic effect than skyrin due to the presence of an additional phenolic hydroxyl group at C-4, which increases oxidative reactive oxygen species generation. This is the first report from P. pinophilum secreating these cytotoxic/proapoptotic secondary metabolites.
authors: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Mytre
authors: M
authors: Koul
authors: Natural Product Chemistry, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Samdarshi
authors: S
authors: Meena
authors: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Ashok
authors: A
authors: Kumar
authors: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Parduman Raj
authors: PR
authors: Sharma
authors: Natural Product Chemistry, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Venugopal
authors: V
authors: Singamaneni
authors: Microbial Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Syed
authors: S
authors: Riyaz-Ul-Hassan
authors: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Abid
authors: A
authors: Hamid
authors: Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Asha
authors: A
authors: Chaubey
authors: Natural Product Chemistry, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Anil
authors: A
authors: Prabhakar
authors: Natural Product Chemistry, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Prasoon
authors: P
authors: Gupta
authors: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
authors: Shashank
authors: S
authors: Singh
mesh_terms: D000880:Anthraquinones / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D027782:Chive / Q000382:microbiology; D004271:DNA, Fungal; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010190:Pancreatic Neoplasms; D010407:Penicillium / Q000737:chemistry* / Q000302:isolation & purification; D017382:Reactive Oxygen Species / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D000880:Anthraquinones; D000970:Antineoplastic Agents; D004271:DNA, Fungal; D017382:Reactive Oxygen Species