BCL2L13 attenuation links impaired mitophagy to epithelial plasticity and anoikis tolerance in lung adenocarcinoma

BCL2L13 is a mitochondrial BCL2-family protein linked to mitophagy and ceramide metabolism, but its role in NSCLC metastatic plasticity remains unclear. Human lung cancer Tissue Microarray (TMA) and matched patient specimens showed subtype- and site dependent BCL2L13 expression, with higher cytoplasmic/granular staining in primary NSCLC and reduced, heterogeneous staining in lymph-node metastases, most evident in adenocarcinoma and squamous-cell carcinoma. Because Epithelial-mesenchymal transition (EMT) and anoikis resistance are central requirements for metastatic dissemination, this primary to node attenuation provided the rationale to test BCL2L13 knockdown and overexpression in metastasis-relevant NSCLC models. In A549 and LLC cell liness, TGF beta 1 induced coordinated mitophagy and EMT with mitochondrial enrichment of BCL2L13. BCL2L13 knockdown impaired TGF beta 1 and carbonyl cyanide m chlorophenyl hydrazone (CCCP) associated mitophagy, reducing LC3 beta mitochondria colocalization, TOMM20 LAMP1 overlap and mitochondrial LC3 II/p62/TOMM20 turnover; BNIP3 and NIX redistribution did not compensate. BCL2L13 loss enhanced EMT-marker switching and migration, whereas overexpression partially opposed these changes. During detachment, BCL2L13 knockdown reduced anoikis-associated apoptosis despite preserved mitochondrial recruitment of BAX/BAK/BNIP3/NIX, altered BID processing, non parallel caspase activity and shifted FAK phosphorylation. Pharmacological autophagy modulation did not reverse this anoikis phenotype. Lipidomics identified adhesion-state-dependent ceramide synthases CerS2/CerS6-linked sphingolipid remodeling: BCL2L13 knockdown increased C24 linked sphingolipid species in attached cells but reduced C16/C24 ceramide-related profiles during anoikis. These findings identify BCL2L13 downregulation as a metastasis-associated mitochondrial–lipid state that limits mitophagic quality control while favoring EMT and detachment survival in NSCLC adenocarcinoma.