Treatment with 188Re Reduces Cancerous Phenotype in Liver Cancer Cells

Abstract

Background Recurrence in hepatocellular carcinoma (HCC) after conventional treatments is a big challenge. Despite the promising progress in advanced targeted therapies, HCC is the fourth leading cause of cancer death worldwide. Radionuclide therapy could be an effective targeted approach to address this concern. Rhenium-188 (188Re) is a β-emitting radionuclide that can be used in clinic for apoptosis induction and inhibit cell proliferation. Although adherent cell cultures are efficient and reliable, the lack of appropriate cell-cell and cell-ECM contact exists. It has been demonstrated that three-dimensional organotypic human cancer models are suitable alternatives. Methods Conventional adherent culture and 3D constructs of Huh7 or HepG2 hepatoma cell lines cultured on liver extracellular matrix (ECM) were treated by different doses of 188 Rhenium Perrhenate (188ReO4). To evaluate cell viability, live/dead assay carried out. The flow-cytometric assay, qRT-PCR, western bolting, colony formation assay, and immunofluorescence (IF) studies were performed to investigate the therapeutic effect of 188ReO4. Subsequently, the tumor formation ability of 188ReO4-treated Huh7 cells was evaluated in animal model. Results According to viability assay and live/dead staining, the number of dead cells in Huh7 and HepG2 lines were significantly increased compared to untreated control groups. Data obtained from Annexin/PI showed that Huh7 and HepG2 cells showed typical apoptotic changes after treatment with 188ReO4. Quantitative RT-PCR and western blotting data also supported that 188ReO4 treatment can induce apoptosis. Furthermore, cell cycle arrest observed in G2 phase after exposure to effective dose of 188ReO4 in Huh7 cells. Colony formation assay confirmed growth suppression in Huh7 and HepG2 cells post exposure. The IF also displayed proliferation inhibition in the 188ReO4 treated cells on 3D scaffolds of liver extracellular matrix (LEM). In 2D culture, PI3-AKT signaling pathway remained unchanged whereas, in the 3D condition it was activated. Treated Huh7 cells with effective dose of 188ReO4 lose their tumor formation ability in nude mice compare to the control group. Conclusion The results supported that 188ReO4 could induce apoptosis and cell cycle arrest and inhibit tumor formation capacity in HCC cells.

Clinical Efficacy of Subgingivally Delivered Punica Granatum Chip and Gel in Management of Chronic Periodontitis Patients

“…13 Various investigations done on both Gram positive and Gram negative non oral bacteria using ethanolic, water, methanolic and acetone extract of pomegranate have shown strong antimicrobial activity. 14 A hydroalcoholic extract of pomegranate fruit was investigated for antibacterial effect on dental plaque microorganisms and was found to be effective against Staphylococcus, Streptococcus, Klebsiella, and Proteus species, as well as Escherichia coli. Most likely ellagitannin and punicalagin are fractions responsible for pomegranate's antibacterial activity.…”