Datkhile Kailas D, Patil Satish R, Durgawale Pratik P, Patil Madhavi N, Hinge Dilip D, Jagdale Nilam J, Deshmukh Vinit N, More Ashwini L
Department of Molecular Biology and Genetics, Krishna Institute of Medical Sciences "Deemed to be University", Taluka-Karad, Dist-Satara, Malkapur, Maharashtra, Pin-415 539, India.
J Genet Eng Biotechnol. 2021 Jan 14;19(1):9. doi: 10.1186/s43141-020-00113-y.
Nanomedicine has evolved as precision medicine in novel therapeutic approach of cancer management. The present study investigated the efficacy of biogenic gold nanoparticles synthesized using Argemone mexicana L. aqueous extract (AM-AuNPs) against the human colon cancer cell line, HCT-15.
Biosynthesis of AM-AuNPs was determined by ultraviolet-visible spectroscopy and further characterized by transmission electron microscopy, X-ray diffraction, and Fourier transition infrared spectroscopy analysis. The cytotoxic activity of AM-AuNPs was assessed by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, whereas genotoxicity was evaluated by the DNA fragmentation assay. The expression of apoptosis regulatory genes such as p53 and caspase-3 was explored through semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and western blotting to evidence apoptotic cell death in HCT-15 cells. Biogenic AM-AuNPs inhibited cell proliferation in HCT-15 cell line with a half maximal inhibitory concentration (IC) of 20.53 μg/mL at 24 h and 12.03 μg/mL at 48 h of exposure. The altered cell morphology and increased apoptosis due to AM-AuNPs were also evidenced through nuclear DNA fragmentation and upregulated expression of p53 and caspase-3 in HCT-15 cells.
The AM-AuNPs may exert antiproliferative and genotoxic effects on HCT-15 cells by cell growth suppression and induction of apoptosis mediated by activation of p53 and caspase-3 genes.
纳米医学已发展成为癌症治疗新方法中的精准医学。本研究调查了使用墨西哥刺蓟(Argemone mexicana L.)水提取物合成的生物源金纳米颗粒(AM-AuNPs)对人结肠癌细胞系HCT-15的疗效。
通过紫外可见光谱法测定AM-AuNPs的生物合成,并通过透射电子显微镜、X射线衍射和傅里叶变换红外光谱分析进一步表征。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法评估AM-AuNPs的细胞毒性活性,而通过DNA片段化测定评估遗传毒性。通过半定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法探索凋亡调节基因如p53和半胱天冬酶-3的表达,以证明HCT-15细胞中的凋亡细胞死亡。生物源AM-AuNPs抑制HCT-15细胞系中的细胞增殖,在暴露24小时时半数最大抑制浓度(IC)为20.53μg/mL,在48小时时为12.03μg/mL。通过核DNA片段化以及HCT-15细胞中p53和半胱天冬酶-3表达上调,也证明了AM-AuNPs导致细胞形态改变和凋亡增加。
AM-AuNPs可能通过抑制细胞生长和激活p53和半胱天冬酶-3基因介导的凋亡对HCT-15细胞发挥抗增殖和遗传毒性作用。