Low‐intensity pulsed ultrasound affects growth, differentiation, migration, and epithelial‐to‐mesenchymal transition of colorectal cancer cells

Abstract

Ultrasound (US) offers potentially important opportunities from a therapeutic point of view. Thus, the study of the biological effects of US on cancer cells is important to understand the consequences of these changes on the malignant phenotype. This study aimed to investigate the effects of low‐intensity ultrasound (LIPUS) on the phenotype of colorectal cancer cell lines. Cell proliferation was evaluated by viability test and by evaluation of pERK expression, while cell motility using the scratch test. Cell differentiation was evaluated assessing alkaline phosphatase activity. Epithelial mesenchymal transition was assessed by analyzing the expression of Vimentin and E‐Cadherin. Release and uptake of extracellular vesicles (EVs) were evaluated by flow cytometry. LIPUS effects on the organization of cytoskeleton were analyzed by confocal microscopy and by evaluation of Rho GTPase expression. No alterations in vitality and clonogenicity were observed when the intermediate (0.4 MPa) and the lowest (0.035 MPa) acoustic intensities were administered while the treatment with high intensity (1 MPa) induced a reduction of both cell viability and clonogenicity in both cell lines in a frequency‐dependent manner. LIPUS promoted the differentiation of colon cancer cells, affected epithelial‐to‐mesenchymal transition, promoted the closure of a wound as well as increased the release of EVs compared with untreated cells. LIPUS‐induced increase in cell motility was likely due to a Rho GTPase‐dependent mechanism. Overall, the results obtained warrant further studies on the potential combined effect of LIPUS with differentiating agents and on their potential use in a clinical setting.

Mechanotransduction in tumor dynamics modeling

“…The proposed model considers the growth and migration of a poroelastic tumor, and the selection of strategic parameters based on the feedback with our experimental results. This allows us to address the possible challenges related to the prediction, controllability, and guidance of new experiments, while effectively avoiding effects associated with cavitation, cytodisruption and standing waves [Mittelstein et al, 2020, Heyden and Ortiz, 2016, Lin et al, 2022, Lucchetti et al, 2020, Prentice et al, 2005. Regarding the complex interplay between LIUS and cancer dynamics, our study confirm that ultrasound does not directly affect healthy cell proliferation and production.…”

“…Recent studies have provided compelling evidence of the therapeutic effectiveness of low-intensity ultrasound. Low-intensity ultrasound (LIUS) and its pulsed version (LIPUS), have been proposed to impact cancer cells by two main mechanisms: i) selectively resonating the right diameter cells under the name of oncotripsy, which lies on destroying the cytoskeleton via cavitation [Mittelstein et al, 2020, Heyden and Ortiz, 2016, Lin et al, 2022, Lucchetti et al, 2020, Prentice et al, 2005, and ii) triggered response produced via mechanotransduction signaling pathways [Lin et al, 2022, Lucchetti et al, 2020, Katiyar et al, 2020, Carina et al, 2018, González et al, 2023, Singh et al, 2021, Tijore et al, 2020, Na et al, 2008, Geiger et al, 2009, Vogel, 2006, Broders-Bondon et al, 2018.…”

Anti-Cancer Role and Therapeutic Potential of Extracellular Vesicles

“…In contrast, there are common sites of cancer metastasis and sites that are specific to the cancer type. In such complicated mechanisms of metastasis, EVs contribute to EMT [ 152 , 153 , 154 ], migration [ 63 , 155 , 156 , 157 ], metastasis niche formation [ 20 , 158 , 159 ], metastasis promotion [ 158 ], and the tumor microenvironment [ 160 ].…”

“…It has been suggested that the microenvironment of cancer contributes to cancer progression through education by EVs. EMT is a feature of cancer metastasis in which epithelial cells transition from the epithelial state to the mesenchymal state, and EVs affecting EMT of cancer cells may lead to cancer metastasis [ 152 , 153 , 154 ]. The migration/invasion step in cancer metastasis is important for migrating to other organs [ 62 , 67 , 68 , 165 , 166 ].…”