评价土壤芽孢杆菌属分离的次生代谢产物的抗菌和抗癌特性，重点关注两株地衣芽孢杆菌和苏云金芽孢杆菌。

Evaluation of antibacterial and anticancer properties of secondary metabolites isolated from soil Bacillus spp focusing on two strains of Bacillus licheniformis and Bacillus siamensis.

机构信息

Department of Microbiology, Faculty of Basic Sciences, Kazerun Branch, Islamic Azad University, Kazerun, Iran.

Department of Microbiology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran.

出版信息

BMC Mol Cell Biol. 2024 Oct 4;25(1):21. doi: 10.1186/s12860-024-00517-5.

DOI:10.1186/s12860-024-00517-5

PMID:39367339

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11451124/

Abstract

BACKGROUND

Bacillus strains are well recognized for their inherent production of bioactive compounds that exhibit antibacterial and anticancer properties. This study aims to evaluate the antimicrobial and anticancer effects of the secondary metabolite isolated from Bacillus licheniformis and Bacillus siamensis strain.

MATERIAL AND METHOD

We developed and purified a new soil-derived Bacillus strain to study its metabolites on cancer cells and bacteria. After evaluating the antimicrobial effects of the selected strains' secondary metabolites by well diffusion, growth conditions and temperature optimised using liquid-liquid extraction, secondary metabolites isolated, and active compounds identified using GC-MS. Evaluation of PC-3 and HPrEpC cytotoxicity. AV/PI staining and comet assay assessed necrosis and apoptosis. Real-time PCR measured apoptotic gene expression. Finally, the scratch test measured cell movement.

RESULTS

Bacillus strain metabolites exhibit dual-purpose antimicrobial and anticancer properties. Bacillus licheniformis isolate 56 and S2-G12 isolate 60 demonstrated the greatest antibacterial activity. Among all Bacillus isolates, isolates 56 (Bacillus licheniformis) and 60 (Bacillus siamensis strain) had the highest antibacterial activity. Crude extracts obtained from strains 56 and 60 decreased PC-3 cell viability in a dose-dependent manner. At 200 µg/mL, the survival rate of cells treated with strain 56 and 60 crude extract was 23% and 25%, respectively (p < 0.001). The treatment of PC-3 cells with strains 56 and 60 crude extract led to considerable apoptosis (46.2% and 50.09%, respectively) compared to the control group. After treatment with the crude extract from strains 56 and 60 at an IC concentration, a significant number of PC-3 cells showed comet formation, indicating DNA fragmentation. Metabolites extracted from strain 56 and 60 enhanced caspase 3, caspase 8, and Bax genes expression and reduced Bcl-2 expression (p < 0.001). Cell migration was also prevented.

CONCLUSION

Our findings show that the secondary metabolites of B. licheniformis and B. siamensis have antibiotic and anticancer properties. However in vivo studies are necessary to confirm these findings.

摘要

背景

芽孢杆菌因其固有产生具有抗菌和抗癌特性的生物活性化合物而广为人知。本研究旨在评估从地衣芽孢杆菌和暹罗芽孢杆菌中分离的次级代谢产物的抗菌和抗癌作用。

材料与方法

我们开发并纯化了一种新的土壤衍生芽孢杆菌菌株，以研究其对癌细胞和细菌的代谢产物。通过平板扩散法评估所选菌株次级代谢物的抗菌效果，优化液体-液体萃取的生长条件和温度，分离次级代谢产物，并使用 GC-MS 鉴定活性化合物。评估 PC-3 和 HPrEpC 的细胞毒性。AV/PI 染色和彗星试验评估坏死和凋亡。实时 PCR 测量凋亡基因表达。最后，划痕试验测量细胞运动。

结果

芽孢杆菌代谢产物具有双重抗菌和抗癌特性。地衣芽孢杆菌分离株 56 和 S2-G12 分离株 60 表现出最强的抗菌活性。在所有芽孢杆菌分离株中，分离株 56（地衣芽孢杆菌）和 60（暹罗芽孢杆菌）的抗菌活性最高。56 株（地衣芽孢杆菌）和 60 株（暹罗芽孢杆菌）菌株获得的粗提取物以剂量依赖性方式降低 PC-3 细胞活力。在 200μg/mL 时，用 56 株和 60 株粗提取物处理的细胞存活率分别为 23%和 25%（p<0.001）。与对照组相比，用 56 株和 60 株粗提取物处理 PC-3 细胞导致显著的细胞凋亡（分别为 46.2%和 50.09%）。在用 56 株和 60 株菌株的粗提取物在 IC 浓度下处理后，大量 PC-3 细胞显示彗星形成，表明 DNA 片段化。从 56 株和 60 株菌株中提取的代谢物增强了 caspase 3、caspase 8 和 Bax 基因的表达，并降低了 Bcl-2 基因的表达（p<0.001）。细胞迁移也被阻止。