Abstract
Biosynthetic silver nanoparticles (AgNPs), specifically formed using medicinal plant extracts, have recently exhibited a remarkable therapeutic effect due to their anticancer potential. Here, we synthesized AgNPs using an aqueous extract of Ginkgo biloba leaves and evaluated its activity against cervical cancer (CCa) and the related molecular mechanisms. The physiochemical properties of the AgNPs were measured by ultraviolet-visible spectrophotometry, nanometre particle size analyzer and transmission electron microscopy. The AgNPs effects on cell proliferation and apoptosis were investigated through MTT, MTS, and colony formation assay; Hoechst 33258 staining; and flow cytometry. The intracellular ROS and oxidative stress levels were assessed using the appropriate commercial kits. Apoptosis-related protein levels were determined by western blotting. We prepared a series of different sized ginkgo extract synthesized AgNPs (GB-AgNPs), and the smallest mean particle size was 40.2 ± 1.2 nm with low polydispersity (0.091 ± 0.011), zeta potential values showed -34.56 mV. Compared to the controls, the GB-AgNP treatment inhibited the cell proliferation and induced the apoptosis of HeLa and SiHa cells. In addition, GB-AgNP treatment led to markedly increased levels of intracellular ROS, the release of cytochrome c (Cyt C) from mitochondria into the cytosol and the cleavage of caspase -9 and -3 in both CCa cell lines. Moreover, NAC, an ROS scavenger, eliminated the effect of GB-AgNPs on the HeLa and SiHa cells. This study reveals that GB-AgNPs suppresses cancer cell proliferation and induces apoptosis by upregulating intracellular ROS generation and inducing the activation of the caspase-dependent mitochondrial apoptotic pathway in CCa cells. Thus, GB-AgNPs may be a potential alternative drug for CCa therapy.
Highlights
In the past several decades, cancer has ranked as the second leading cause of death in worldwide [1]
We found that gingko leaf extract-based synthesis of silver nanoparticles (GB-AgNPs) exhibited good inhibitory effects on cell proliferation and induced the apoptosis of cervical cancer (CCa) cells
An annexin V-FITC/PI apoptosis detection Kit, reactive oxygen species (ROS) assay kit, malondialdehyde (MDA) assay kit, glutathione peroxidase (GSH-Px) assay kit and superoxide dismutase (SOD) assay kit were purchased from KeyGen Biotech (Beijing, CN).The primary antibodies against b-actin, Bax, Bcl-2, Caspase-9, Caspase-3, cytochrome c (Cyt C), and Vadc 1 were purchased from Cell Signaling (Massachusetts, USA)
Summary
In the past several decades, cancer has ranked as the second leading cause of death in worldwide [1]. Cervical cancer (CCa) is becoming more prevalent, in terms of both incidence and mortality, which are ranked fourth, globally [2]. Nanoparticles (NPs) are new materials with dimension between approximately 1 and 100 nm in at least one dimension and in as many as three dimensions [5]. Due to their physiochemical properties and characteristics, NPs exhibit therapeutic potential for numerous diseases [6]. Synthetic biologically active AgNPs have exhibited extensive therapeutic potential with broad spectrum antibacterial, antifungal, anticancer, anti-inflammatory, wound healing, antioxidative, and anti-diabetic activity [7,8,9]
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