Resveratrol-Induced Temporal Variation in the Mechanical Properties of MCF-7 Breast Cancer Cells Investigated by Atomic Force Microscopy

Abstract

Atomic force microscopy (AFM) combined with fluorescence microscopy has been used to quantify cytomechanical modifications induced by resveratrol (at a fixed concentration of 50 µM) in a breast cancer cell line (MCF-7) upon temporal variation. Cell indentation methodology has been utilized to determine simultaneous variations of Young’s modulus, the maximum adhesion force, and tether formation, thereby determining cell motility and adhesiveness. Effects of treatment were measured at several time-points (0–6 h, 24 h, and 48 h); longer exposures resulted in cell death. Our results demonstrated that AFM can be efficiently used as a diagnostic tool to monitor irreversible morpho/nano-mechanical changes in cancer cells during the early steps of drug treatment.

Time- and Zinc-Related Changes in Biomechanical Properties of Human Colorectal Cancer Cells Examined by Atomic Force Microscopy

“…HT-29-MTX cells incubated under ZD conditions for 96 h showed remarkable changes, with a drop in the maximum adhesion of around 60% down to 240 pN. This lower adhesion was similar to that from HT-29-MTX cultivated in the presence of zinc as well as ZD HT-29, and was comparable to other cell lines measured previously under similar conditions (MCF7, Caco-2) [ 18 , 24 ]. The mean adhesion force of ZA HT-29 cultivated for 96 h, however, was twice as high as that for ZA HT-29-MTX at the same time-point.…”

“…This demands a comprehensive collection of data on the biomechanics of single (cancerous) cells, which has been mainly generated by studying different in vitro cultured carcinoma cell lines [8]. In fact, these cell lines are often used to examine concentration and time-dependent exposure of cancerous cells to different pharmacological or bioactive compounds via AFM [18,19]. Biomechanical properties have already been studied for various tissues [20], among them the gastrointestinal tract as well as the colon [21], though this technique has been predominantly applied to study non-pathological tissues [22,23], and less to differentiate between healthy and deteriorated, abnormal tissue from colon biopsies.…”

Nucleotides-Induced Changes in the Mechanical Properties of Living Endothelial Cells and Astrocytes, Analyzed by Atomic Force Microscopy

“…AFM measurements were accomplished with a Nanowizard III instrument (JPK Instruments, Berlin, Germany) incorporating a Cell Hesion module (which allows the extension of the piezo Z range to 100 µm), mounted on an inverted optical microscope (Axio Observer Z1, Zeiss, Jena, Germany). The AFM was operated in force-spectroscopy mode, applying constant setpoint (1.5 nN) and loading rate (5 µm/s), as previously described [ 58 ]. These values were chosen to avoid an influence on the cell mechanics outcome, as recently described by Weber et al [ 41 ].…”

Biomechanics of Cell Membrane

“…On the same line, Jagoba Iturri et al [ 2 ] employed Atomic force microscopy (AFM) combined with fluorescence microscopy, to quantify cytomechanical modifications induced by resveratrol (at a fixed concentration of 50 μM) in a breast cancer cell line (MCF-7), upon temporal variation. They quantified Young’s modulus, the maximum adhesion force, and tether formation, and thereby, they determined important insight into cell motility and adhesiveness.…”

“…Moreover, time dependence fits well into three distinct levels (short incubation, 24, and 48 h) according to the impact observed. Maintenance of a drug incubation for longer periods (48 h) induced a gradual loss of properties that concludes in cell death [ 2 ]. These results confirmed the validity of the AFM technique as a decisive tool to detect irreversible transformations at the nanoscale level, that might affect normal cell functioning, or even their malignancy, as in the case of tumoral cells during the early steps of drug treatment.…”