Nadumane Varalakshmi K, Venkat Prerana, Pal Anamika, Dharod H, Shukla Megha, Prashanthi K
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University, #18/3, 9 Main, III Block, Jayanagar, Bangalore-560 011, India.
Indian J Pharm Sci. 2013 Sep;75(5):507-14.
This study aims at the isolation of filamentous fungi, extraction of metabolites, and evaluation of the cytotoxic properties on HeLa cells and normal human lymphocytes. We isolated fungi from the soil by serial dilution method. One of the isolates was chosen and identified as Aspergillus ochraceus Wilhelm (Trichocomaceae) by standard techniques. The metabolites were extracted using methanol. Different concentrations of the extract were evaluated for their potential anticancer activity on HeLa cells by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide assay and the safety of the extract was checked on normal human lymphocytes. The extract was purified by chromatographic techniques like thin-layer chromatography and high-performance liquid chromatography, and subjected to mass spectrometric analysis. The extract showed significant cytotoxic potential on HeLa cells at low concentrations with a half maximal inhibitory concentration value of <50 μg/ml. The extract gave 10 fractions by thin layer chromatography, and fraction B had higher toxicity than the rest. This fraction gave a single peak by high-performance liquid chromatography and had a mass-to-charge ratio of 905.65, which did not match any of the earlier known fungal metabolites or metabolites from other strains of A. ochraceus. The metabolite from A. ochraceus is alkaloid in nature, cytotoxic to HeLa cells, and appears to be a novel with anticancer potentials, which could be explored further for characterization of the active component.
本研究旨在分离丝状真菌、提取代谢产物，并评估其对HeLa细胞和正常人淋巴细胞的细胞毒性特性。我们采用连续稀释法从土壤中分离真菌。选择其中一株分离物，并通过标准技术鉴定为赭曲霉Wilhelm（曲霉科）。使用甲醇提取代谢产物。通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑法评估不同浓度提取物对HeLa细胞的潜在抗癌活性，并检测提取物对正常人淋巴细胞的安全性。提取物通过薄层色谱和高效液相色谱等色谱技术进行纯化，并进行质谱分析。提取物在低浓度下对HeLa细胞显示出显著的细胞毒性潜力，半数最大抑制浓度值<50μg/ml。提取物通过薄层色谱得到10个馏分，B馏分的毒性高于其他馏分。该馏分通过高效液相色谱得到一个单峰，质荷比为905.65，与任何先前已知的真菌代谢产物或其他赭曲霉菌株的代谢产物均不匹配。赭曲霉的代谢产物本质上是生物碱，对HeLa细胞具有细胞毒性，似乎是一种具有抗癌潜力的新型物质，可进一步探索其活性成分的特性。
