内生真菌是一种被充分证实的生物活性化合物库，这些化合物具有药用价值，因此在生物医学领域中发挥重要作用。本研究旨在从药用植物龙骨花叶子的内生真菌-球果茎病菌（Colletotrichum gloeosporioides）的乙酸乙酯提取物中鉴定出生物活性抗癌化合物。采用抗氧化活性引导分离结合串联液相色谱-四极杆飞行时间质谱、傅里叶变换红外光谱、飞行时间质谱和核磁共振技术，已经鉴定出脂肪酸酰胺化合物N-(2-羟乙基)十六烷酰胺（巴洛米托醇胺，PEA）。体外分子对接分析表明，PEA可以潜在地与调控细胞凋亡的蛋白质，包括BAX、BCL-2、P21和P53等有活性的结合。进一步验证用体外研究证明，PEA可以抑制乳腺癌细胞MDA-MB-231和MCF-7的增殖，改变细胞核形态，减弱伤口愈合能力。PEA通过上调细胞周期阻滞基因（P21）、抑制肿瘤（P53）、促凋亡（BAX、CASPASE-8 和 FADD）基因下调抗凋亡基因BCL-2诱导细胞凋亡。报告了Caspase-3活性的上调。这是第一次报道从C. gloeosporioides中分离到PEA，显示其对人类乳腺癌细胞具有抗癌活性，因此具有未来治疗潜力。©2023年国际生物化学和分子生物学联盟。

Endophytic fungi are a well-established reservoir of bioactive compounds that are pharmaceutically valuable and therefore, contribute significantly to the biomedical field. The present study aims to identify the bioactive anticancer compound from ethyl acetate extract of fungal endophyte, Colletotrichum gloeosporioides associated with the leaf of the medicinal plant Oroxylum indicum. The fatty acid amide compound N-(2-Hydroxyethyl)hexadecanamide (Palmitoylethanolamide; PEA) was identified using antioxidant activity-guided fractionation assisted with tandem liquid chromatography coupled with quadrupole time of flight mass spectrometry, Fourier transform-infrared spectroscopy, time-of-flight mass spectrometry, and nuclear magnetic resonance. In-Silico molecular docking analysis showed that PEA potentially docked to the active sites of apoptosis-inducing proteins including BAX, BCL-2, P21, and P53. Further validation was done using in vitro study that showed PEA inhibitsthe proliferation, alters nuclear morphology and attenuates the wound closure ability of MDA-MB-231 and MCF-7 cells. PEA induces apoptosis via upregulating cell-cycle arrest (P21), tumor suppression (P53), pro-apoptotic (BAX, CASPASE-8, and FADD) genes, and downregulating anti-apoptotic gene BCL-2. The upregulation of the active form of Caspase-3 was also reported. This is the first-ever report for the isolation of PEA from C. gloeosporioides with anticancer activity against human breast cancer cells and therefore holds great potential for future therapeutics.© 2023 International Union of Biochemistry and Molecular Biology.