Pourramezan Zahra, Oloomi Mana, Kasra-Kermanshahi Rouha
Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran.
Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran.
Int J Prev Med. 2020 Sep 3;11:132. doi: 10.4103/ijpvm.IJPVM_307_19. eCollection 2020.
It has been proven that probiotic bacteria have inhibitory effects on human cancer cell lines. The aim of this study is to isolate and characterize the antioxidant probiotic and determine the possible anticancer activities of the selected strain.
One of the strain isolated from camel doogh sample showed the high antioxidant activity by using of different methods such as resistance to hydrogen peroxide, hydroxyl radical and superoxide anions. The antioxidant strain was characterized by sequencing of 16S rRNA V2-V3 regions and the 16S-23S intergenic spacer (ITS). The methanol extract of this strain supernatant was fractionated using thin layer chromatography (TLC) and antioxidant activity of fractions was detected by 0.1% of DPPH through TLC-DPPH bioautography. anticancer activity of each fraction was investigated by using MTT and flow cytometry methods.
According to the phylogenetic results, the antioxidant strain was closely related to strain E91 (Accession No. EF536365). After fractionation and anti-proliferation assessments of strain AG12a extracellular materials, one of the antioxidant fraction (F4) showed maximum DPPH radical scavenging activity (IC of 535.27 μg/mL). MTT assay of the F4 fraction demonstrated cytotoxic activity against Caco-2 with the IC50 value of 299.05 μg/mL. The cell death activity of the fraction was confirmed by flow cytometry with 30.925.
In this study, the anticancer and apoptotic properties of against Caco-2 cell line was reported for the first time. The isolated bioactive fraction from the extracellular methanol extract needs to be further investigated in human studies of cancer therapy.
已证实益生菌对人类癌细胞系具有抑制作用。本研究的目的是分离和鉴定抗氧化益生菌,并确定所选菌株可能的抗癌活性。
从骆驼酸奶样品中分离出的一株菌株,通过对过氧化氢、羟基自由基和超氧阴离子的抗性等不同方法显示出高抗氧化活性。通过对16S rRNA V2-V3区域和16S-23S基因间隔区(ITS)进行测序来鉴定抗氧化菌株。该菌株上清液的甲醇提取物用薄层色谱(TLC)进行分离,通过TLC-DPPH生物自显影法检测各馏分的抗氧化活性。通过MTT和流式细胞术方法研究各馏分的抗癌活性。
根据系统发育结果,该抗氧化菌株与菌株E91(登录号EF536365)密切相关。对菌株AG12a细胞外物质进行分离和抗增殖评估后,其中一个抗氧化馏分(F4)显示出最大的DPPH自由基清除活性(IC为535.27μg/mL)。F4馏分的MTT分析表明对Caco-2具有细胞毒性活性,IC50值为299.05μg/mL。通过流式细胞术以30.925证实了该馏分的细胞死亡活性。
本研究首次报道了该菌株对Caco-2细胞系的抗癌和凋亡特性。从细胞外甲醇提取物中分离出的生物活性馏分需要在癌症治疗的人体研究中进一步研究。