Chronically Established ρ0 Cancer Cells Exhibit Shared Stemness-Associated and Aggressive Phenotypes Across Multiple Cancer Cell Lines.

Chronically established mitochondrial DNA-depleted (ρ0) cancer cells may exhibit phenotypic changes beyond metabolic dysfunction; however, whether such changes are shared across different cancer types remains unclear. In this study, we generated chronically established ρ0 models from pancreatic, lung, and breast cancer cell lines and examined their common phenotypic features. Compared with their parental wild-type counterparts, the ρ0 cell lines exhibited increased proliferation, chemoresistance-related features, sphere-forming ability, wound closure, invasion-related phenotypes, and angiogenic activity, together with common morphological alterations. These changes were accompanied by VDAC1 upregulation and cytoskeletal remodeling. Pharmacologic treatment with VBIT-4 attenuated several of these phenotypes, whereas its effects on chemoresistance-related responses varied across cell lines. Collectively, these findings suggest that chronically established ρ0 cancer cells exhibit shared stemness-associated and aggressive phenotypes across multiple cancer cell lines, accompanied by VDAC1 upregulation and cytoskeletal remodeling.