Raman and ATR-FTIR Spectroscopic Profiling Coupled with Gene Expression to Monitor Physiopathological Dynamics in 3D HepG2 Liver Cancer Spheroids †

The integration of micro-Raman and ATR-FTIR spectroscopies with qRT-PCR gene expression analysis provides a multimodal strategy for profiling three-dimensional hepatocellular carcinoma (HCC) spheroids. This approach enables temporal monitoring of tumor growth, metabolic adaptation, and microenvironmental remodeling. Gene expression revealed an early proliferative phase (PCNA, KI-67, AFP), followed by activation of hypoxia- and stress-associated pathways (HIF1α, SLC2A1/GLUT1, LDHA) and modulation of extracellular matrix (ECM) components (SPARC, FN1) alongside the pro-apoptotic regulator BBC3. In parallel, Raman and ATR-FTIR spectroscopies non-invasively detected biochemical variations related to oxidative stress, lipid accumulation, apoptosis, and ECM remodeling. The observed correlations between spectral signatures and molecular markers demonstrate that these label-free spectroscopic fingerprints are reliable indicators of physiopathological processes. Our research integrates molecular spectroscopy with transcriptomics to provide an innovative and comprehensive approach for a quick and first evidence of tumor progression.