Zina Hamada A, Kalaba Mohamed H, Shaban Abdelghany S, Elrefaey Ahmed A, Mahdy Hesham M, Haikal Abdullah
Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo, 11884, Egypt.
Department of Pharmacognosy, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt.
Sci Rep. 2025 Sep 12;15(1):32472. doi: 10.1038/s41598-025-18565-y.
Cancer remains a leading cause of death worldwide, necessitating the discovery of novel therapeutic agents with improved efficacy and selectivity. This study investigated the anticancer potential of Aspergillus niger strain AW17 isolated from honeybees, a previously unexplored ecological niche for anticancer drug discovery. The fungal isolate was characterized through morphological, microscopic, and molecular methods, confirming its identity as Aspergillus niger. Chemical profiling using GC-MS and UPLC-MS/MS revealed a complex composition including high oleic acid content (28.88%) and unique compounds like pachymic acid (18.93%), rarely associated with A. niger. The extract's anticancer activity was evaluated against colorectal (Caco-2), liver (HepG2), breast (MCF-7), and lung (A549) cancer cell lines using MTT assay, confocal microscopy with AO/PI staining, cell cycle analysis, and apoptosis detection. Results demonstrated remarkable selective cytotoxicity of the Aspergillus niger extract against cancer cell lines, following the sensitivity hierarchy HepG2 (IC = 5.22 µg/ml) > Caco-2 (26.78 µg/ml) > A549 (34.18 µg/ml) > MCF-7 (55.91 µg/ml), while sparing normal cell lines significantly, with IC values of 1454.7 µg/ml for WI-38 and 668.3 µg/ml for HFB-4, indicating high cancer cell selectivity. Confocal microscopy confirmed membrane integrity loss as a primary cytotoxic mechanism. Cell cycle analysis revealed cell-type specific arrest patterns, with Caco-2 and HepG2 exhibiting G1 arrest, while MCF-7 and A549 showed S phase accumulation. Flow cytometry indicated distinct death mechanisms, with Caco-2 undergoing early apoptosis and substantial necrosis, HepG2 showing predominant late apoptosis, and A549 displaying primary necrotic death. These findings highlight the potential of honeybee-associated fungi as sources of novel anticancer compounds with remarkable selectivity, providing new avenues for targeted cancer therapy development and demonstrating the value of exploring specialized ecological niches in natural product discovery.
癌症仍是全球主要死因，因此需要发现疗效和选择性更佳的新型治疗药物。本研究调查了从蜜蜂中分离出的黑曲霉菌株AW17的抗癌潜力，蜜蜂是一个此前未被探索过的用于抗癌药物发现的生态位。通过形态学、显微镜和分子方法对该真菌分离株进行了鉴定，确认其为黑曲霉。使用气相色谱 - 质谱联用仪（GC-MS）和超高效液相色谱 - 串联质谱仪（UPLC-MS/MS）进行的化学分析表明，其成分复杂，包括高含量的油酸（28.88%）以及如茯苓酸（18.93%）等独特化合物，这些化合物很少与黑曲霉相关联。使用MTT法、AO/PI染色共聚焦显微镜、细胞周期分析和凋亡检测等方法，对该提取物针对结肠癌细胞系（Caco-2）、肝癌细胞系（HepG2）、乳腺癌细胞系（MCF-7）和肺癌细胞系（A549）的抗癌活性进行了评估。结果表明，黑曲霉提取物对癌细胞系具有显著的选择性细胞毒性，敏感性顺序为HepG2（IC = 5.22 μg/ml）＞Caco-2（26.78 μg/ml）＞A549（34.18 μg/ml）＞MCF-7（55.91 μg/ml），同时对正常细胞系有显著保护作用，WI-38的IC值为1454.7 μg/ml，HFB-4的IC值为668.3 μg/ml，表明对癌细胞具有高选择性。共聚焦显微镜证实膜完整性丧失是主要的细胞毒性机制。细胞周期分析揭示了细胞类型特异性的停滞模式，Caco-2和HepG2表现为G1期停滞，而MCF-7和A549表现为S期积累。流式细胞术表明存在不同的死亡机制，Caco-2经历早期凋亡和大量坏死，HepG2主要表现为晚期凋亡，A549表现为原发性坏死死亡。这些发现突出了蜜蜂相关真菌作为具有显著选择性的新型抗癌化合物来源的潜力，为靶向癌症治疗的发展提供了新途径，并证明了在天然产物发现中探索特殊生态位的价值。
