Apigenin restores apoptotic pathways and induces G2/M arrest via modulation of DNA methylation and expression of pro and anti-apoptotic genes in HeLa cells

Apoptosis resistance is a characteristic of cervical cancer advancement, caused by the loss of tumor suppressor activity and continuous activation of pro-survival pathways. This study examines apigenin, a dietary flavonoid, as a regulator of proliferation and apoptotic signaling in HeLa cervical cancer cells. in silico analysis reported that Apigenin had a robust binding affinity for CDK1, CDK2, XIAP, and cIAP, indicating competitive suppression of their natural ligand interactions. In vitro studies revealed that treatment of HeLa cells with Apigenin (24 & 48 H, 1-175 µM) resulted in a dose- and time-dependent reduction in cell viability, mitochondrial membrane depolarization, G2/M phase arrest, and an increase in apoptotic cell populations. These phenotypic alterations were followed by the overexpression of pro-apoptotic genes and tumor suppressors (DIABLO, BAX, CASP3, CASP8, p53, DAPK1) and the downregulation of anti-apoptotic genes (BCL2, XIAP, Survivin), which was consistent at the protein level. Methylation sequencing across CpG contexts demonstrated that differential DNA methylation coincided with transcriptional changes, with alterations in CpG island methylation rather than non-CpG contexts exhibiting the most significant correlation with the modified expression of apoptotic and anti-apoptotic genes. These findings establish CpG methylation as a crucial epigenetic mechanism driving apigenin-induced transcriptional reprogramming, positioning apigenin as a dual-function agent that induces apoptosis and modulates the cervical cancer epigenome, with implications for enhanced cancer management strategies.