Laboratory of Animal Genetic Engineering and Stem Cell Biology, Department of Animal Biotechnology, Jeju National University, Jeju, Jeju-Do 690-756, Republic of Korea.
Department of Biotechnology, Division of Research and Development, Lovely Professional University, Punjab 144411, India.
Mater Sci Eng C Mater Biol Appl. 2019 Apr;97:166-176. doi: 10.1016/j.msec.2018.11.055. Epub 2018 Nov 28.
BRM270 is the most leading phytochemical extract that possesses potent anticancer properties. A major challenge associated with this drug is its low bioavailability and thus requires high dosages for cancer treatment. Here, we report the novel nano-synthesis of phyto-composite, BRM270 for the first time by mechanical milling method with specific modifications for enhanced cytotoxicity against HepG2 human hepatoma cancer cells. Unlike free BRM270 and other phytomedicines, BRM270 nanoparticles (BRM270 NPs) are well-dispersed and small sized (23 to 70 nm) which is believed to greatly enhanced cellular uptake. Furthermore, the acidic tumor microenvironment attracts BRM270 NPs enhancing targeted therapy while leaving normal cells less affected. The comparative cytotoxicity analysis using MTT assay among the three treatment groups, such as free BRM270, BRM270 NPs, and doxorubicin demonstrated that BRM270 NPs induced greater cytotoxicity against HepG2 cells with an effective drug concentration of 12 μg/ml. From FACS analysis, we observed an apoptotic cell death of 44.4% at BRM270 NPs treated cells while only 12.5% found in the free BRM270 treated cells. Further, the comparative relative expression profiling of the candidate genes were showed significant (p < 0.05) down-regulation of IL6, BCL2, p53, and MMP9 in the BRM270 NPs treated cells, compared to the free BRM270 and doxorubicin. Indeed, the genes, CASPASE 9 and BAX have shown significant (p < 0.05) upregulation in cells treated with BRM270 NPs as compared to counter treatment groups. The investigation of the signal pathways and protein-protein network associations were also carried out to elucidate the functional insights underlying anti-cancer potential of BRM270 NPs in HepG2 cells. Taken together, our findings demonstrated that these uniquely engineered BRM270 NPs effectively enter into the cancer cells due to its acidic microenvironment thereby inducing apoptosis and regulate the cell-proliferation in-vitro at extremely low dosages.
BRM270 是最主要的植物化学提取物，具有强大的抗癌特性。这种药物的一个主要挑战是其生物利用度低，因此需要高剂量用于癌症治疗。在这里，我们首次通过机械研磨法报告了植物复合材料 BRM270 的新型纳米合成，该方法具有特定的修饰，可增强对 HepG2 人肝癌癌细胞的细胞毒性。与游离 BRM270 和其他植物药不同，BRM270 纳米颗粒（BRM270 NPs）分散良好且尺寸较小（23 至 70nm），这被认为极大地增强了细胞摄取。此外，酸性肿瘤微环境吸引 BRM270 NPs 增强靶向治疗，同时使正常细胞受到的影响较小。MTT 测定法对三种治疗组（游离 BRM270、BRM270 NPs 和阿霉素）的比较细胞毒性分析表明，BRM270 NPs 在有效药物浓度为 12μg/ml 时对 HepG2 细胞诱导更强的细胞毒性。从 FACS 分析中，我们观察到在 BRM270 NPs 处理的细胞中发生了 44.4%的凋亡细胞死亡，而在游离 BRM270 处理的细胞中仅发现了 12.5%。此外，候选基因的相对表达谱分析表明，与游离 BRM270 和阿霉素相比，BRM270 NPs 处理的细胞中 IL6、BCL2、p53 和 MMP9 的表达显著（p<0.05）下调。事实上，与对照治疗组相比，细胞中 CASPASE 9 和 BAX 的基因表达显著（p<0.05）上调。还进行了信号通路和蛋白质-蛋白质网络关联的研究，以阐明 BRM270 NPs 在 HepG2 细胞中抗癌潜力的功能见解。总之，我们的研究结果表明，这些独特设计的 BRM270 NPs 由于其酸性微环境而有效地进入癌细胞，从而诱导细胞凋亡，并以极低的剂量在体外调节细胞增殖。
