In this comprehensive study, a robust methodology to investigate the regulatory effects of ENO1 and DDIT4 gene silencing on cell proliferation and apoptosis, underscoring their critical roles in cancer biology. Genes linked to programmed cell death (PCD) across 12 distinct PCD patterns were collated from multiple sources, resulting in a final set of 1078 genes analyzed for their involvement in apoptosis, pyroptosis, and other cell death pathways. Functional enrichment analyses conducted through DAVID and Kyoto encyclopedia of genes and genomes (KEGG) pathways yielded insights into the molecular functions and biological processes influenced by these genes. Key analyses included the use of ssGSEA to calculate cell death scores and unsupervised clustering to identify patterns in acute myeloid leukemia (AML) samples, which revealed significant associations with patient prognosis. A cell death index (CDI) through the least absolute shrinkage and selection operator (LASSO) Cox regression model, validated across different cohorts, and demonstrated predictive value for patient outcomes. Experimental assessments further confirmed the role of ENO1 and DDIT4 in modulating apoptosis, where gene silencing demonstrated significant effects on cell death and proliferation markers like BCL2, Caspase-3, FAS, and BAX. These findings were supported by flow cytometry assays, which quantitatively assessed apoptosis in U937 and MV411 cells, thereby reinforcing the genes’ roles in cancer pathology. Furthermore, the study investigated the interface between CDI and immune cell infiltration, demonstrating that CDI correlates with immune landscape alterations within the tumor microenvironment (TME), thus also suggesting potential implications for immunotherapy responsiveness. Collectively, the study provides a multi-dimensional view of how specific genes influence cell death mechanisms and their broader implications in cancer therapy and prognosis.