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Mechanical Properties of Colorectal Cancer Cells Determined by Dynamic Atomic Force Microscopy: A Novel Biomarker.

ABSTRACT: Colorectal cancer (CRC) has been addressed in the framework of molecular, cellular biology, and biochemical traits. A new approach to studying CRC is focused on the relationship between biochemical pathways and biophysical cues, which may contribute to disease understanding and therapy development. Herein, we investigated the mechanical properties of CRC cells, namely, HCT116, HCT15, and SW620, using static and dynamic methodologies by atomic force microscopy (AFM). The static method quantifies Young's modulus; the dynamic method allows the determination of elasticity, viscosity, and fluidity. AFM results were correlated with confocal laser scanning microscopy and cell migration assay data. The SW620 metastatic cells presented the highest Young's and storage moduli, with a defined cortical actin ring with distributed F-actin filaments, scarce vinculin expression, abundant total focal adhesions (FAK), and no filopodia formation, which could explain the lessened migratory behavior. In contrast, HCT15 cells presented lower Young's and storage moduli, high cortical tubulin, less cortical F-actin and less FAK, and more filopodia formation, probably explaining the higher migratory behavior. HCT116 cells presented Young's and storage moduli values in between the other cell lines, high cortical F-actin expression, intermediate levels of total FAK, and abundant filopodia formation, possibly explaining the highest migratory behavior.

SUBMITTER: Bras MM

PROVIDER: S-EPMC9600571 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature

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Mechanical Properties of Colorectal Cancer Cells Determined by Dynamic Atomic Force Microscopy: A Novel Biomarker.

Brás M Manuela MM Cruz Tânia B TB Maia André F AF Oliveira Maria José MJ Sousa Susana R SR Granja Pedro L PL Radmacher Manfred M

Cancers 20221015 20

Colorectal cancer (CRC) has been addressed in the framework of molecular, cellular biology, and biochemical traits. A new approach to studying CRC is focused on the relationship between biochemical pathways and biophysical cues, which may contribute to disease understanding and therapy development. Herein, we investigated the mechanical properties of CRC cells, namely, HCT116, HCT15, and SW620, using static and dynamic methodologies by atomic force microscopy (AFM). The static method quantifies ...[more]

PMID: 36291838

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Project description:BackgroundLignin is a complex polymer which inhibits the enzymatic conversion of cellulose to glucose in lignocellulose biomass for biofuel production. Cellulase enzymes irreversibly bind to lignin, deactivating the enzyme and lowering the overall activity of the hydrolyzing reaction solution. Within this study, atomic force microscopy (AFM) is used to compare the adhesion forces between cellulase and lignin with the forces between cellulase and cellulose, and to study the moiety groups involved in binding of cellulase to lignin.ResultsTrichoderma reesei, ATCC 26921, a commercial cellulase system, was immobilized onto silicon wafers and used as a substrate to measure forces involved in cellulase non-productive binding to lignin. Attraction forces between cellulase and lignin, and between cellulase and cellulose were compared using kraft lignin- and hydroxypropyl cellulose-coated tips with the immobilized cellulase substrate. The measured adhesion forces between kraft lignin and cellulase were on average 45% higher than forces between hydroxypropyl cellulose and cellulase. Specialized AFM tips with hydrophobic, -OH, and -COOH chemical characteristics were used with immobilized cellulase to represent hydrophobic, H-bonding, and charge-charge interactions, respectively. Forces between hydrophobic tips and cellulase were on average 43% and 13% higher than forces between cellulase with tips exhibiting OH and COOH groups, respectively. A strong attractive force during the AFM tip approach to the immobilized cellulase was observed with the hydrophobic tip.ConclusionsThis work shows that there is a greater overall attraction between kraft lignin and cellulase than between hydroxypropyl cellulose and cellulase, which may have implications during the enzymatic reaction process. Furthermore, hydrophobic interactions appear to be the dominating attraction force in cellulase binding to lignin, while a number of other interactions may establish the irreversible binding.

Dataset Information

Mechanical Properties of Colorectal Cancer Cells Determined by Dynamic Atomic Force Microscopy: A Novel Biomarker.

Publications

Mechanical Properties of Colorectal Cancer Cells Determined by Dynamic Atomic Force Microscopy: A Novel Biomarker.

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