Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-11, Iran.
Arch Microbiol. 2022 Sep 23;204(10):641. doi: 10.1007/s00203-022-03258-1.
Actinomycetes are filamentous bacteria and the residents of the soil, prone to produce bioactive metabolites. This research aimed to isolate, classify, and investigate the anticancer properties of Actinomycetes secondary metabolites from various saline soils of Qom province. Actinomycetes isolates were molecularly recognized by 16SrRNA gene sequencing after the PCR procedure. The A549 cell line was then exposed to bacterial metabolites to find their cytotoxicity by MTT assay and their capacity to cause apoptosis by Flow cytometry. The expression levels of the bax and bcl-2 genes were determined using Real-time PCR. Bacterial metabolites were distinct by HPLC and GC-MS assays. Sequencing identified three novel Actinomycetes strains, Streptomyces griseoflavus, Streptomyces calvus, and Kitasatospora phosalacineus. The IC doses of bacterial metabolites were discovered equal to 1337, 2619, and 4874 µg/ml, respectively. Flow cytometric assay revealed that their secondary metabolites were capable of inducing apoptosis in A549 cells by 25%, 14.5%, and 7.58%, respectively. Real-time PCR findings displayed that the bax gene expression in A549 cells treated with S. griseoflavus and S. calvus, comparatively increased (P < 0.0008, P < 0.00056). The expression of the bcl-2 gene was significantly reduced in cells treated with S. griseoflavus and K. phosalacineus (P < 0.0006, P < 0.0004). The findings of this analysis showed the presence of new isolates in a soil sample from Qom province which can produce new anticancer agents and can be considered appropriate candidates for further research to employ as anticancer drugs.
放线菌是丝状细菌,也是土壤居民,容易产生生物活性代谢物。本研究旨在从库姆省各种盐土中分离、分类和研究放线菌次生代谢物的抗癌特性。在 PCR 程序后,通过 16SrRNA 基因测序对放线菌分离物进行分子识别。然后将 A549 细胞系暴露于细菌代谢物中,通过 MTT 测定法检测其细胞毒性,并通过流式细胞术检测其诱导细胞凋亡的能力。使用实时 PCR 测定 bax 和 bcl-2 基因的表达水平。通过 HPLC 和 GC-MS 分析鉴定细菌代谢物的特征。测序鉴定出三种新型放线菌菌株,即灰色链霉菌、草生链霉菌和游动放线菌。细菌代谢物的 IC 剂量分别为 1337、2619 和 4874μg/ml。流式细胞术检测表明,它们的次生代谢物能够分别诱导 A549 细胞凋亡 25%、14.5%和 7.58%。实时 PCR 结果显示,用 S. griseoflavus 和 S. calvus 处理的 A549 细胞中 bax 基因的表达显著增加(P<0.0008,P<0.00056)。用 S. griseoflavus 和 K. phosalacineus 处理的细胞中 bcl-2 基因的表达显著降低(P<0.0006,P<0.0004)。本分析结果表明,在库姆省土壤样本中存在新的分离物,它们可以产生新的抗癌剂,可被视为进一步研究的合适候选药物,用于开发抗癌药物。