Analysis of the A549 cell line affected by anticancer bioactive compounds of Actinomycetes isolated from saline soils.
Amin KhoshakhlaghSaeid AbrounSeyyed Soheil AghaeiMohammad SoleimaniMohammad Reza ZolfaghariPublished in: Archives of microbiology (2022)
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 50 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.
Keyphrases
- induced apoptosis
- cell cycle arrest
- ms ms
- real time pcr
- endoplasmic reticulum stress
- cell death
- gene expression
- oxidative stress
- flow cytometry
- genome wide
- poor prognosis
- single cell
- high throughput
- heavy metals
- south africa
- escherichia coli
- signaling pathway
- dna methylation
- mass spectrometry
- genome wide identification
- binding protein
- genetic diversity
- risk assessment
- human health
- simultaneous determination
- high performance liquid chromatography
- cell proliferation
- tissue engineering