Investigation of the anticancer effects of sorafenib, spirulina platensis ex-tract and combined applications on hepatocellular cancer cells

Hepatocellular carcinoma (HCC) is one of the most prevalent primary liver tumors worldwide and is associated with a high mortality rate. Sorafenib, a multikinase inhibitor used in the systemic treatment of advanced-stage HCC, is known for its ability to suppress cell proliferation, inhibit angiogenesis, and induce apoptosis. However, its limited efficacy when used alone, alongside its side effect profile and the potential for resistance development, has necessitated the investigation of complementary therapeutic agents. In this context, the present study aimed to explore, for the first time, the anticancer effects of the natural microalga Spirulina platensis in combination with Sorafenib on hepatocellular carcinoma cells. The originality of this thesis lies in the experimental assessment of this novel therapeutic combination. The experimental design involved the use of the human hepatocellular carcinoma cell line (HepG2) as a cancer model and the human umbilical vein endothelial cell line (HUVEC) as a healthy control model. Cells were treated for 24 and 48 hours with varying concentrations of Sorafenib (1–15 µM) and Spirulina platensis extract (1–100 µg/mL). The cytotoxic, apoptotic, anti-inflammatory, and genotoxic effects of these treatments were evaluated through assessments of cell viability, gene and protein expression, and DNA damage. Cell viability was measured using the MTT assay, while quantitative real-time PCR (qRT-PCR) was employed to analyze the expression of apoptosis- and inflammation-related genes including BAX, CASP3, BCL-2, and TNF-α. ELISA assays were used to assess the corresponding protein levels, and the Comet Assay was conducted to determine the extent of DNA damage. The findings demonstrated that the combination of 5 µM Sorafenib and 100 µg/mL Spirulina extract significantly reduced the viability of HepG2 cells while maintaining relatively higher survival rates in HUVEC cells. Molecular analyses revealed increased expression of pro-apoptotic genes (BAX and CASP3), downregulation of the anti-apoptotic gene BCL-2, and modulation of TNF-α expression consistent with an anti-inflammatory response. Protein expression data supported the gene-level findings, and DNA damage was notably increased in high-dose combination groups. These results suggest that the co-administration of Spirulina platensis with Sorafenib may elicit a synergistic anticancer effect in HCC treatment. However, the observed genotoxicity at higher doses underscores the importance of dose optimization. This study highlights the potential of naturally derived bioactive compounds as supportive agents in modern targeted cancer therapies and contributes valuable preliminary data for future preclinical and clinical investigations.