Ghate Nikhil Baban, Chaudhuri Dipankar, Sarkar Rhitajit, Sajem Albert L, Panja Sourav, Rout Jayashree, Mandal Nripendranath
Division of Molecular Medicine, Bose Institute, Kolkata, India.
Department of Ecology and Environmental Science, Assam University, Silchar, Assam, India.
PLoS One. 2013 Dec 16;8(12):e82293. doi: 10.1371/journal.pone.0082293. eCollection 2013.
This report highlights the phytochemical analysis, antioxidant potential and anticancer activity against breast carcinoma of 70% methanolic extract of lichen, Parmotrema reticulatum (PRME). Phytochemical analysis of PRME confirms the presence of various phytoconstituents like alkaloids, carbohydrates, flavonoids, glycosides, phenols, saponins, tannins, anthraquinones, and ascorbic acid; among which alkaloids, phenols and flavonoids are found in abundant amount. High performance liquid chromatography (HPLC) analysis of PRME revealed the presence of catechin, purpurin, tannic acid and reserpine. Antioxidant activity was evaluated by nine separate methods. PRME showed excellent hydroxyl and hypochlorous radical scavenging as well as moderate DPPH, superoxide, singlet oxygen, nitric oxide and peroxynitrite scavenging activity. Cytotoxicity of PRME was tested against breast carcinoma (MCF-7), lung carcinoma (A549) and normal lung fibroblast (WI-38) using WST-1 method. PRME was found cytotoxic against MCF-7 cells with an IC50 value 130.03 ± 3.11 µg/ml while negligible cytotoxicity was observed on A549 and WI-38 cells. Further flow cytometric study showed that PRME halted the MCF-7 cells in S and G2/M phases and induces apoptosis in dose as well as time dependent manner. Cell cycle arrest was associated with downregulation of cyclin B1, Cdk-2 and Cdc25C as well as slight decrease in the expression of Cdk-1 and cyclin A1 with subsequent upregulation of p53 and p21. Moreover PRME induced Bax and inhibited Bcl-2 expression, which results in increasing Bax/Bcl-2 ratio and activation of caspase cascade. This ultimately leads to PARP degradation and induces apoptosis in MCF-7 cells. It can be hypothesised from the current study that the antioxidant and anticancer potential of the PRME may reside in the phytoconstitutents present in it and therefore, PRME may be used as a possible source of natural antioxidant that may be developed to an anticancer agent.
本报告重点介绍了地衣网脉梅衣70%甲醇提取物(PRME)的植物化学分析、抗氧化潜力及对乳腺癌的抗癌活性。PRME的植物化学分析证实了其含有多种植物成分,如生物碱、碳水化合物、黄酮类、糖苷、酚类、皂苷、单宁、蒽醌和抗坏血酸;其中生物碱、酚类和黄酮类含量丰富。PRME的高效液相色谱(HPLC)分析显示存在儿茶素、紫茜素、单宁酸和利血平。通过九种不同方法评估了抗氧化活性。PRME表现出优异的羟基和次氯酸根自由基清除能力,以及中等程度的二苯基苦味酰基自由基(DPPH)清除能力、超氧阴离子清除能力、单线态氧清除能力、一氧化氮清除能力和过氧亚硝酸盐清除能力。使用WST - 1法检测了PRME对乳腺癌(MCF - 7)、肺癌(A549)和正常肺成纤维细胞(WI - 38)的细胞毒性。发现PRME对MCF - 7细胞具有细胞毒性,IC50值为130.03±3.11μg/ml,而在A549和WI - 38细胞上观察到的细胞毒性可忽略不计。进一步的流式细胞术研究表明,PRME使MCF - 7细胞停滞在S期和G2/M期,并以剂量和时间依赖性方式诱导细胞凋亡。细胞周期停滞与细胞周期蛋白B1、细胞周期蛋白依赖性激酶2(Cdk - 2)和细胞分裂周期蛋白25C(Cdc25C)的下调以及细胞周期蛋白依赖性激酶1(Cdk - 1)和细胞周期蛋白A1表达的轻微降低相关,随后p53和p21上调。此外,PRME诱导Bax表达并抑制Bcl - 2表达,这导致Bax/Bcl - 2比值增加和半胱天冬酶级联反应激活。这最终导致聚(ADP - 核糖)聚合酶(PARP)降解并诱导MCF - 7细胞凋亡。从当前研究可以推测,PRME的抗氧化和抗癌潜力可能存在于其中所含的植物成分中,因此,PRME可能作为一种天然抗氧化剂的潜在来源,有望开发成为一种抗癌药物。