Investigation of cell membrane protein signatures in two-dimensional and three-dimensional NSCLC models under normoxic and hypoxic conditions

Lung cancer, one of the leading causes of cancer-related deaths in man and woman. Cancer stem cells (CSCs) play an important role in tumor recurrence, progression, and therapy resistance. Therefore, targeting CSCs plays a vital role in the diagnosis and treatment of lung cancers. In this study, 3D (three-dimensional) non-small cell lung cancer (NSCLC) tumoroids were generated and exposed to a hypoxic microenvironment to induce CSC development. Enriched cell membrane proteins were isolated from tumoroids grown under hypoxic and normoxic conditions. The enriched cell membrane protein lysate obtained was analyzed by proteomic methods. In total, 2225 proteins were identified under all conditions. A significant portion of these identified proteins (approximately 40%) were localized in the cell membrane. Proteomic analysis were identified CSC-related proteins (such as DLL1, NOTCH1 TSPAN5, THBS1, CALR, PLXNB2, ABCA5, NOTCH2NLB, etc.) in normoxic and hypoxic conditions. As a result, these proteins obtained under normoxic and hypoxic microenvironmental conditions in this study present potential biomarker candidates for the diagnosis and treatment of NSCLC.