Dose-dependent effects of wheat germ oil and vitamin E on apoptosis and gene expression in breast and cervical cancer cell lines with single-cell RNA-seq profiling

The search for natural sources has raised hopes for a natural metabolite with fewer side effects in the treatment of cancers. Therefore, this study investigated the therapeutic effects of different doses of wheat germ oil (WGO) and α-tocopherol (vitamin E) on MCF-7 and HeLa cell lines through the expression of apoptosis genes and Single-cell RNA seq analysis (scRNA-seq). Cells were treated with WGO (15 µg/ml, 7.5 µg/ml and 3.75 µg/ml) and α-tocopherol concentrations for 14 days after plating in cell culture plates. After that, apoptotic cells were examined using flow cytometry with Annexin V-FITC and PI staining. Cell cycle analysis was performed by flow cytometry, expression analysis of BAX, BCL-2 and P53 genes by real-time PCR (RT-PCR), and scRNA-seq of cervical cancer cells were evaluated. Concentrations of 15 µg/ml and 7.5 µg/ml WGO and α-tocopherol led to an increase in necrotic and apoptotic cells and a decrease in viable cells in MCF-7 and HeLa cell lines (p < 0.05). While the concentration of 3.75 µg/ml did not show a significant effect (p > 0.05). In addition, gene expression analysis showed that treatment with different concentrations of WGO and α-tocopherol significantly reduced BCL-2 gene expression (p < 0.05) and increased P53 and BAX gene expression in both cell lines (p < 0.05). The study highlights the beneficial effects of high doses of WGO (15 and 7.5 µg/mL) and α-tocopherol in MCF-7 and HeLa cell lines by examining cell viability, apoptosis and necrosis, and apoptotic pathway genes.