Evaluating the Effects of Nano-niosome Co-loaded with Artemisinin and Metformin on the Survival and Proliferation of A549 Lung Cancer Cells

Abstract

Metformin and Artemisinin are herbal medicines that have been proven to have anti-cancer properties. Studies have shown that these two drugs inhibit the growth and proliferation of cancer cells by inhibiting different signaling pathways such as mTOR, MAPK, AMPK and NF-kB. According to the findings of previous studies, the loading of anticancer drugs in nanoparticles such as niosomes increases the anticancer effects of drugs due to the increase in solubility and appropriate release of drugs at the target site. The purpose of the upcoming study is to evaluate the anticancer effects of metformin and artemisinin drugs alone and loaded in nisome nanoparticles and compare the anticancer effects of the drugs in free form and loaded inside nisome nanoparticles on A549 cell line. Material and Method: PEGylated niosome nanoparticles containing metformin and artemisinin were synthesized and their physicochemical properties were investigated using scanning electron microscope (SEM), infrared spectroscopy (FT-IR), and DLS. The release rate of drugs loaded from nanoparticles was measured by placing the drug-containing nanoparticle in the release medium. In order to investigate the toxicity effect of PEGylated niosome containing metformin and Artemisinin on A549 cell line, MTT test was performed. Apoptosis test was performed to evaluate the effect of PEGylated niosomes containing metformin and Artemisinin on the programmed death of A549 cells. Investigating the effect of drugs loaded inside PEGylated niosomes on stopping the cell cycle was done by Cell Cycle test. Finally, the expression level of hTERT, caspase-7, caspase-3, BAX, BCL-2, and cyclin D1 genes was investigated in the presence of pegylated niosomes containing metformin and Artemisinin. Findings: Microscopic findings and FT-IR results showed successful drug loading. The results of MTT showed significant cell arrest compared to the control group. Drug toxicity evaluation showed that metformin and artemisinin loaded in PEGylated-niosome had more toxic effects compared to the pure state in a time- and dose-dependent manner. Although the separate treatment of Metformin and Artemisinin stopped the growth and proliferation of A549 cells, the simultaneous treatment of these two drugs increased the inhibitory effects on the growth and proliferation of cells in the culture medium. The results of Real-Time PCR showed that metformin and artemisinin in a free and loaded form in nanoparticles inhibit hTERT, BCL-2 and cyclin D1 genes expression and increased the expression of caspase-7, caspase3 and BAX expression. The simultaneous treatment of these two drugs causes a further decrease in the expression of these two genes. The results of the apoptosis test showed that cell death in the presence of PEGylated-niosome containing metformin and artemisinin is of the apoptotic type.