Programmed cell death (PCD) is a ubiquitous phenomenon in eukaryotic cells but is less understood in bacteria, where limited instances such as endospore formation in Bacillus subtilis have been experienced earlier. In our study, we investigated the induction of PCD in Pseudomonas aeruginosa triggered by colistin exposure which leads to generation of the increased number of persister cells under biofilm embeded conditions. Interestingly, when colistin was combined with nitroxoline, a transition from apoptosis to necrosis was noted, rendering the cells more vulnerable to damage. Gene expression analyses revealed that overproduction of recA, typically involved in bacterial DNA repair pathways, facilitated PCD induction under colistin stress. The primary mode of damage was mediated by reactive oxygen species (ROS), which caused genomic DNA destruction, protein damage, and bacterial membrane disruption through lipid peroxidation. As a possible effect of PCD, a higher number of persister cells was evident in the multicellular organization such as biofilm in the colistin-treated sample. Furthermore, in the presence of the colistin-nitroxoline combination, severe membrane damage was observed due to cation chelation, leading to cell death from significant osmotic shock. Understanding these mechanisms of antibiotic-mediated killing is crucial for selecting effective treatments in clinical settings.