Abstract

Cervical cancer remains a global health burden disproportionately affecting low- and middle-income countries (LMICs) due to inaccessible prevention strategies, screening, and treatment. In this exploratory, in vitro proof-of-concept study, we investigated the potential synergistic antiproliferative effect of combining ivermectin and ascorbic acid against the cervical cancer model (HeLa) cell line. Antiproliferative effects of ascorbic acid and ivermectin were evaluated in HeLa cell lines using the MTT assay. Using experimentally calculated half maximal inhibitory concentration (IC50) values for ivermectin and ascorbic acid, the two drugs were combined, and their interactions were analyzed using Compusyn software. Computational prediction was performed using network pharmacology and molecular docking studies. Gene expression analysis was also performed to provide functional validation of putative targets by assessing their deregulation at the transcriptional level. IC50 values of ivermectin and ascorbic acid on HeLa cells were 9.65 μM and 4.58 mM, respectively. Analysis using Compusyn software for the interaction between ascorbic acid and ivermectin showed an additive effect near the IC50 and synergy at higher concentrations (>IC50). Network pharmacology and molecular docking predicted putative genes and pathways for the combination treatment. Further, gene expression analysis on selected genes showed upregulation of TP53 and downregulation of STAT3, BCL2, HIF1α, and TNF. Overall, this exploratory proof-of-concept study demonstrates dose-dependent antiproliferative activity between ivermectin and ascorbic acid, though with weak synergism.

Key words: Ascorbic acid, Cervical cancer, Drug repurposing, HeLa cells, Ivermectin