Phytochemical Composition, Antibacterial and Anticancer Activities of Trifolium Cherleri Extract on Lung Cancer Cell Line (A549) and Analysis of Caspase 3 and Caspase 9 Apoptosis Genes Expression



Mirzaie A¹ ; Kashtali AB¹ ; Sahebjamee H² ; Noorbazargan H³ ; Rahmati H⁴ ; Sadat Shandiz SA⁵
Source: Tehran University Medical Journal Published:2017

Abstract

Background: Medicinal plants have been identified and used from prehistoric times and these plants make many chemical compounds for biological functions. Trifolium cherleri is an herbaceous species belonging to the family of the Fabaceae to Africa, Eurasia and Australia. T. cherleri is an important member of the Fabaceae family that is well-known herbal medicine in Iran. The aim of this study was to investigate the phytochemical composition, antibacterial and anti-cancer activities of T. cherleri extract. Methods: This experimental study was performed in Islamic Azad University, from December 2016 to February 2017. At first, the phytochemical constituents of T. cherleri extract were determined using gas chromatography-mass spectrometry (GCMS) method. Subsequently, the antibacterial activity of the extract was evaluated against some gram positive and negative pathogenic bacteria included Staphylococcus aureus ATCC 25923, Streptococcus pyogenes ATCC 19615, Salmonella enteritidis ATCC 13076 and Listeria monocytogenes ATCC 35152 via minimum inhibitory concentration (MIC) method. Moreover, anticancer potential of extract was examined by colorimetric MTT assay toward lung cancer (A549) cell line. Then, the evaluation of caspase 3 and 9 apoptosis gene expression was determined using Real-Time Polymerase Chain Reaction (Real-Time PCR) technique. Moreover, the Real-Time PCR was performed using relative quantitative method. Results: The phytochemical analyses of T. cherleri extract showed the 20 major components and the most frequent component was belonged to hexadecanoic acid, ethyl ester (20.7%) and 2-Pentadecanone, 6,10,14-trimethyl (19.9%). The extract had maximum antibacterial effects against Staphylococcus aureus and Streptococcus pyogenes. There was a dose dependent increase in the cytotoxicity effect of extract against A549 cancer cell. Moreover, the Real-Time PCR results indicated that the caspase 3 and caspase 9 gene expression was significantly up-regulated 2.57±0.27 (P<0.05), and 3.3±0.46 (P<0.05), respectively. Conclusion: The results of this study showed that the T. cherleri extract had significant anti-bacterial and anti-cancer effects and it appear that the extract has potential uses for pharmaceutical industries. Moreover, it could be considered as a promising source for novel drug compounds, but more studies are needed. © 2017, Tehran University of Medical Sciences. All rights reserved.