Abstract
We used three label-free minimally invasive methods to characterize individual cells derived from primary and secondary tumours from the same patient, and of the same type – colorectal. Raman spectroscopy distinguished cells by their biochemical 'fingerprint' in a vibrational spectrum with 100% accuracy, and revealed that the primary cell line contains more lipids and alpha-helix proteins, whereas the secondary cell line contains more porphyrins and beta-sheet proteins. Stimulated Raman scattering (SRS) microscopy distinguished cells in chemically-specific images of CH2 bonds which revealed lipid droplets in secondary tumour cells. Atomic force microscopy (AFM) was used to distinguish cells with 80% accuracy by measuring their elasticity – secondary tumour cells (SW620) are around 3 times softer than primary ones (SW480). As well as characterizing the physical and biochemical differences between cell lines in vitro, these techniques offer three novel methods which could potentially be used for diagnosis – to assign a tumour as primary or secondary.
Highlights
Metastasis – the spreading of cancer from one organ to another – results in secondary tumours which are responsible for 90% of deaths from cancer.[1]
Cells were plated onto glass-bottom dishes (WPI Fluorodish FD35) for stimulated Raman scattering (SRS) microscopy and atomic force microscopy (AFM), and onto 0.15–0.18 mm thick quartz substrates (SPI supplies 1019T-AB) for Raman spectroscopy
We found the mean values and standard deviations of elasticity to be 1.39 ± 0.71 kPa (SW480 retrace) and 0.834 ± 1.43 kPa (SW620 retrace), using the retraction part of the force–distance curve
Summary
Metastasis – the spreading of cancer from one organ to another – results in secondary tumours which are responsible for 90% of deaths from cancer.[1] To reduce death rates, it is essential to understand metastasis and characterize primary and secondary tumours. Primary tumours possess different genetic profiles[2,3] from secondary tumours, so this is used to clinically diagnose whether a tumour is primary or secondary in nature. Receptor biomarkers are specific to a cancer type,[4] and these biomarkers of a secondary tumour should remain unchanged from the primary source. Some biomarkers may be specific to metastasis.[5] So, in future it may be possible to distinguish primary and secondary tumours with MRI or CT contrast agents, such as nanoparticles coated with such antibodies
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