Evaluation of Ramucirumab as a combinational cancer therapeutic with 5-Aza- citidine in human Hepatoma (HuH-7) cell line

Abstract

The purpose of this study is to explore the impact of Ramucirumab alone and combined with 5-Aza on different cellular targets in human hepatocellular carcinoma cell line (HuH-7). Cells were treated with diverse concentrations of Ramucirumab (50, 100, 150 µg/mL) for 24 h. viability, levels of reactive oxygen species (ROS), antioxidant activities, mitochondrial membrane potential (MMP), DNA fragmentation, and expression of apoptotic genes (Caspases 3, 7, 9, p53, Bax, and Bcl-2) were measured. Results showed that HuH-7 cell viability was reduced in dose-dependent way with Ramucirumab therapy. Both Ramucirumab and 5-Aza elevated ROS, but their combination reduced ROS levels, though still higher than controls. Ramucirumab decreased SOD activity; while 5-Aza increased it. Combined therapy potentially increased SOD and CAT activities. Ramucirumab alone was not responsible to influence GSH levels, but 5-Aza and the combination treatments increased GSH levels. All treatments reduced the JC-1 aggregate/monomer ratio, indicating decreased MMP and potential apoptosis induction. High fragmentated DNA and condensation of chromatin were recorded suggesting apoptosis. Both agents, either as monotherapy or combined therapy, upregulated Caspases 3, 7, and 9, p53, and Bax expression, with variable effects on Bcl-2. In conclusion, Ramucirumab, particularly when co-administered with 5-Aza, effectively reduces viability of cells and prompts apoptosis in HuH-7 cells through increased oxidative stress, disturbance of MMP, and apoptotic pathways activation. The combination treatment shows potential for enhanced therapeutic efficacy against hepatocellular cancer cells.

Keywords

• Ramucirumab
• 5-Aza
• Apoptosis
• Hepatocellular carcinoma
• Oxidative stress