Following a period of disuse owing to the emergence of multidrug-resistant Gram-negative bacteria, colistin has regained global attention as an antibiotic of last resort. The resurgence in its utilization has led to a concurrent increase in acquired resistance, presenting a significant challenge in clinical treatment. Predominantly, resistance mechanisms involve alterations in the lipid A component of the lipopolysaccharide (LPS) structure. These alterations are facilitated by the addition of cationic phosphoethanolamine (pEtN) or 4-amino-L-arabinose, often driven by the upregulation of two-component regulatory systems such as PmrAB and PhoPQ. Structural components of bacteria, such as capsules and efflux pumps, can also play an important role in the resistance mechanism. In addition to these biochemical modifications, structural components of bacteria like capsules and efflux pumps also play crucial roles in mediating resistance. Another significant mechanism is the acquisition of the plasmid-mediated mobilized colistin resistance (mcr) gene, which poses a global health threat due to its ability to transfer between different bacterial genera. Contemporary strategies to combat colistin resistance include the development and use of novel drugs and inhibitors. To devise effective interventions, it is imperative to first elucidate the precise mechanisms of colistin resistance and determine the roles of various contributing factors.