The therapaeutic effects of Apigenin and Ramucirumab on experimentally induced hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver, originating from hepatocytes. Apigenin has antioxidant, anti-inflammatory, antibacterial, and antiviral properties. In addition, apigenin has been shown to have tumor suppressor activity in recent years. Ramucirumab (RAM) is a monoclonal antibody that inhibits vascular endothelial growth factor receptor-2 (VEGFR-2) and is clinically used in cancer treatment. In our study, the effects of Apigenin and Ramucirumab on the levels of STAT-3, iNOS, CIP2A, NF-κB, PI3K, AKT, AFP, PTEN, and VEGFR-2 were investigated in an experimental hepatocellular carcinoma model induced by diethylnitrosamine (DEN). For this study, thirty male Wistar Albino rats were divided into five groups of six: control, sham, DEN, DEN+Apigenin, and DEN+Ramucirumab. At the beginning of the experiment, all groups, except for the control and sham groups, were intraperitoneally (i.p.) injected with DEN (50 mg/kg) once a week for seven weeks. The control group received no treatment, while the sham group received only a 10% DMSO solution during this seven-week period. Following the induction, there was a two-week interval with no treatment for any group. Starting from the 10th week, Apigenin (25 mg/kg) was administered daily, and Ramucirumab (6 mg/kg) was administered three times a week (Monday, Wednesday, and Friday), both dissolved in 10% DMSO and delivered via i.p. injection for a duration of two weeks. At the end of the experiment, blood and liver tissue samples were collected. Plasma and tissue supernatants were evaluated using the ELISA method. The remaining liver tissue samples were stained with hematoxylin-eosin, and the detected lesions were classified histopathologically. According to the findings, the DEN group exhibited increased levels of STAT-3, iNOS, CIP2A, NF-κB, PI3K, AKT, PTEN, VEGFR-2, and AFP in both serum and liver tissue compared to the control group. In the Apigenin treatment group, serum levels of AFP, VEGFR-2, PTEN, and CIP2A were significantly reduced, whereas the decrease in serum levels of AKT, PI3K, NF-κB, STAT-3, and iNOS was not significant. In liver tissue, while levels of AFP, VEGFR-2, PI3K, and NF-κB were significantly decreased, the reduction in the levels of AKT, PTEN, CIP2A, STAT-3 and iNOS did not reach statistical significance. In the Ramucirumab treatment group, a significant decrease was observed in serum levels of AFP, VEGFR-2, PTEN, CIP2A and iNOS, while the reduction in serum levels of AKT, PI3K, NF-κB and STAT-3 was not significant. Although Ramucirumab reduced all parameters in the liver tissue, only the decrease in the STAT-3 level was not statistically significant. Histopathological evaluations demonstrated a strong correlation with the biochemical findings. The DEN group showed lesion areas containing hydropic degeneration, necrosis, and atypical hepatocytes. In contrast, the Apigenin group exhibited rare atypical cells, and no atypical cells were observed in the Ramucirumab group. In conclusion, both Apigenin and Ramucirumab were found to exhibit chemotherapeutic effects; however, the anti-carcinogenic effect of Ramucirumab was more potent. This suggests that the difference may be attributed to the distinct mechanisms of action of the two agents.