Inhibition of Carbonic Anhydrase IX Suppresses Breast Cancer Cell Motility at the Single-Cell Level.

Agne Janoniene,Vilma Petrikaite,Linas Mazutis,Daumantas Matulis

doi:10.3390/ijms222111571

Agne Janoniene, Vilma Petrikaite + Show 2 more

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https://doi.org/10.3390/ijms222111571

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Abstract

Protein Carbonic Anhydrase IX (CA IX), which is expressed in various hypoxic solid tumors in order to maintain proper pH, is also related to cancer cell adhesion, invasion, and metastasis processes. Here, we investigated whether CA IX inhibition by a highly CA IX selective agent benzenesulfonamide VD11-4-2 triggers changes in individual cell motility. We seeded breast cancer cells on an extracellular matrix-coated glass-bottomed dish and in a microfluidic device with a gradient flow of epidermal growth factor (EGF), tracked individual cell movement, calculated their migration speeds, and/or followed movement direction. Our results showed that the inhibitor VD11-4-2 decreased the speed of CA IX positive breast cancer cells by 20–26% while not affecting non-cancerous cell migration. The inhibitor suppressed the cell migration velocity increment and hindered cells from reaching their maximum speed. VD11-4-2 also reduced CA IX, expressing cell movement towards the growth factor as a chemoattractant. Such a single cell-based migration assay enabled the comprehensive investigation of the cell motility and revealed that VD11-4-2 shows the ability to suppress breast cancer cell migration at a lower concentration than previously tested CA IX inhibitors.

Highlights

Breast cancer is the most common type of cancer occurring in women worldwide (WHO)
In vitro studies with breast cancer cell lines have shown that Carbonic Anhydrase IX (CA IX) inhibitors could have the potential to affect cell migration during a scratch assay [11,13,14]
VD11-4-2 was further tested by using the MCF-7 breast cancer line, which is derived from a metastatic site [31]

Summary

Introduction

Breast cancer is the most common type of cancer occurring in women worldwide (WHO). the early stages of breast cancer can be controlled, progressed malignancies or metastases are often associated with poor survival [1]. A wide range of agents acting on different targets is being tested in preclinical studies with the purpose of reducing tumor metastasis, but none of them are approved for clinical use [3] This leads to a search for novel targets that would affect one of the fundamental steps of metastasis. In vitro studies with breast cancer cell lines have shown that CA IX inhibitors (ureido sulfonamides and sulfamates) could have the potential to affect cell migration during a scratch assay [11,13,14] These studies monitored cell migration during the scratch assay, which is based on monitoring how monolayered cells occupy the empty cell culturing dish space, while cell metastasis is more commonly a single cell interaction with extracellular matrix (ECM) processes [15,16]

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