Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana State, India.
Academy of Scientific and Innovative Research (AcSIR), CSIR-HRDC Campus, Kamala Nehru Nagar, Gaziabad 201002, U.P., India.
ACS Appl Bio Mater. 2024 Aug 19;7(8):5057-5075. doi: 10.1021/acsabm.4c00597. Epub 2024 Aug 8.
Melanoma has gained considerable attention due to its high mortality and morbidity rate worldwide. The currently available treatment options are associated with several limitations such as nonspecificity, drug resistance, easy clearance, low efficacy, toxicity-related issues, etc. To this end, nanotechnology has garnered significant attention for the treatment of melanoma. In the present manuscript, we have demonstrated the and anticancer activity of silver nitroprusside nanoparticles (abbreviated as AgNNPs) against melanoma. The AgNNPs exhibit cytotoxicity against B16F10 cells, which has been investigated by several experiments including [methyl H]-thymidine incorporation assay, cell cycle and apoptosis analysis by flow cytometry, and ROS generation through DCFDA, DHE, and DAF2A reagents. Further, the internalization of nanoparticles was determined by ICPOES analysis, while their colocalization was analyzed by confocal microscopy. Additionally, JC-1 staining is performed to examine mitochondrial membrane potential (MMP). Cytoskeleton integrity was observed by phalloidin staining. Expression of different markers (Ki-67, cytochrome c, and E-cadherin) was checked using an immunofluorescence assay. The therapeutic efficacy of AgNNPs has been validated in the melanoma model established by inoculating B16F10 cells into the dorsal right abdomen of C57BL/6J mice. The intraperitoneal administration of AgNNPs reduced melanoma growth and increased the survivability of tumor-bearing mice. The immunofluorescence studies (Ki-67, CD31, and E-cadherin) and TUNEL assay support the inhibitory and apoptotic nature of AgNNPs toward melanoma, respectively. Furthermore, the various signaling pathways and molecular mechanisms involved in anticancer activity are evaluated by Western blot analysis. These findings altogether demonstrate the promising anticancer potential of AgNNPs toward melanoma.
黑色素瘤因其在全球范围内的高死亡率和发病率而引起了相当大的关注。目前可用的治疗选择存在一些局限性，如特异性差、耐药性、易清除、疗效低、毒性相关问题等。为此，纳米技术在治疗黑色素瘤方面引起了广泛关注。在本手稿中，我们展示了银硝普钠纳米颗粒（简称 AgNNPs）对黑色素瘤的 和 抗癌活性。AgNNPs 对 B16F10 细胞表现出细胞毒性，这已通过几种 实验包括 [甲基 H]-胸腺嘧啶掺入测定、细胞周期和凋亡分析通过流式细胞术以及通过 DCFDA、DHE 和 DAF2A 试剂生成 ROS 来研究。此外，通过 ICPOES 分析确定了纳米颗粒的内化，通过共聚焦显微镜分析了它们的共定位。此外，通过 JC-1 染色检查线粒体膜电位 (MMP)。通过鬼笔环肽染色观察细胞骨架完整性。使用免疫荧光测定法检查不同标志物（Ki-67、细胞色素 c 和 E-钙粘蛋白）的表达。通过将 B16F10 细胞接种到 C57BL/6J 小鼠的背部右侧腹部来验证 AgNNPs 的 治疗功效。AgNNPs 的腹腔内给药减少了黑色素瘤的生长并提高了荷瘤小鼠的存活率。免疫荧光研究（Ki-67、CD31 和 E-钙粘蛋白）和 TUNEL 测定分别支持 AgNNPs 对黑色素瘤的抑制和凋亡特性。此外，通过 Western blot 分析评估了涉及抗癌活性的各种信号通路和分子机制。这些发现共同证明了 AgNNPs 对黑色素瘤具有有前途的抗癌潜力。
