Distinct spectral signatures unfold ECM stiffness-triggered biochemical changes in breast cancer cells.

Cancer progression often accompanies the stiffening of extracellular matrix (ECM) in and around the tumor, owing to extra deposition and cross-linking of collagen. Stiff ECM has been linked with poor prognosis and is known to fuel invasion and metastasis, notably in breast cancer. However, the underlying biochemical or metabolic changes and the cognate molecular signatures remain elusive. Here, we explored Raman spectroscopy to unveil the spectral fingerprints of breast cancer cells in response to extracellular mechanical cues. Using stiffness-tuneable hydrogels, we showed that cells grown on stiff ECM displayed morphological changes with high proliferation. We further demonstrated that Raman Spectroscopy, a label-free and non-invasive technique, could provide comprehensive information about the biochemical environment of breast cancer cells in response to varying ECM stiffness. Raman spectroscopic analysis classified the cells into distinct clusters based on principal component-based linear discriminant analysis (PC-LDA). Multivariate curve resolution-alternating least squares (MCR-ALS) analysis indicated that cells cultured on stiff ECM exhibited elevated nucleic acid content and lesser lipids. Interestingly, increased intensity of Raman bands corresponding to cytochrome-c was also observed in stiff ECM conditions, suggesting mitochondrial modulation. The key findings harboured by spectral profiles were also corroborated by transmission electron microscopy, confirming altered metabolic status as reflected by increased mitochondria number and decreased lipid droplets in response to ECM stiffening. Collectively, these findings not only give the spectral signatures for mechanoresponse but also provide the landscape of biochemical changes in response to ECM stiffening.

Serum Raman spectroscopy: Prognostic applications in oral cancers.

BACKGROUND: Loco-regional recurrences attributable to field cancerization and minimal residual cancer, remain prime causes of mortality in oral cancer (OC) subjects. The current study evaluates potential of serum Raman spectroscopy (SRS) to identify recurrence-prone OC subjects. METHODS: Raman spectra of serum from eight healthy subjects (H) and 57 OC subjects (with-recurrence [R], without-recurrence [NR], and with suspicious-lesions [S]), before (BS) and after (AS) surgical excision of tumor were recorded. OC subjects were followed-up for 7-years. RESULTS: DNA and protein alterations were observed in AS sera of all groups. 4-, 3-, and 2-model multivariate analyses were used to stratify BS and AS groups. H spectra were 100% distinguishable from all other groups. AS, R and NR were distinguished with high accuracy (84%) in all models. No stratification (~50%) was observed BS. CONCLUSION: SRS shows potential to identify recurrence prone subjects, post-surgery, using serum collected as early as 1 week after surgery.

Raman Spectroscopy Revealed Cell Passage-Dependent Distinct Biochemical Alterations in Radiation-Resistant Breast Cancer Cells.

Recapitulating radioresistant cell features in pertinent cell line models is essential for deciphering fundamental cellular mechanisms. The limited understanding of passage and cell cycle phases on radioresistant cells revived post-cryopreservation led us to investigate the effect of sub-culturing in parental and radioresistant MCF-7 cells. In this study, the radioresistant cells showed high-intensity nucleic acid and cytochrome bands, which are potentially a radiation-induced spectral marker. Raman spectroscopy data showed dynamic biochemical alterations in revived radioresistant G2/M synchronized cells at early cell passages 1 and 3 with stabilization at a latter cell passage, 5. The study highlights the importance of cell passaging and cell cycle phases in potentially changing the biochemical parameters during in vitro experiments after the revival of radioresistant cells post-cryopreservation.