Abstract
Background
Higher prevalence of hepatocellular carcinoma (HCC) in men is largely attributed to the sex hormones and behavioral risk factors. This study investigated the effects of male (testosterone) and female (estradiol) hormones on Aurora-A/B kinases, GATA-3, and microRNAs (miR-3941, miR-4500, miR-4742) in HepG2 and Huh-7 cell lines.Methods
In this study, two human hepatocellular carcinoma cell lines, HepG2 (with a differentiated phenotype) and Huh-7 (with more aggressive tumorigenic behavior), were treated with the sex hormones β-estradiol and testosterone. Hormonal effects were evaluated using the MTT assay (to assess cytotoxicity), flow cytometry (for cell cycle analysis), RT-qPCR (to quantify the expression of target genes and miRNAs, including Aurora-A, Aurora-B, GATA-3, miR-3941, miR-4500, and miR-4742), and western blotting (to assess protein levels of Aurora-A/B kinases and GATA-3).Results
In HepG2 cells, estradiol and testosterone decreased Aurora-A/B kinase expression and increased GATA3, causing G0/G1 and G2/M cell cycle arrest and reduced DNA synthesis. Conversely, in the more aggressive Huh7 cell line, these hormones accelerated G1-S transition and led to late cell cycle accumulation. Both cell lines exhibited inverse miRNA regulation (miR-3941 and miR-4500 upregulated; miR-4742 downregulated), suggesting a post-transcriptional mechanism affecting cell division genes. Multivariate regression indicated Aurora-A/B independently regulated G0/G1-S transition in HepG2, while temporal factors predominated in Huh7 cell cycle control.Conclusion
A simultaneous comparison of female and male sex hormones in two HCC cell models-one less aggressive (HepG2) and one highly aggressive (Huh-7)-demonstrated that the direction of hormonal effects (inhibitory or stimulatory) depends not on hormone type but on cellular phenotype, and downstream regulatory networks. This duality of response was similar for both hormones within each cell line. This paradox may be attributed to the predominance of distinct signaling pathways-protective in HepG2 versus proliferative in Huh-7 cells.