Assessing the Relationship Between Bladder Cancer cell Contractile Phenotypes and their Invasive Properties

Increased cell contractility is considered a hallmark of highly aggressive tumor cells in several cancers. However, the relationship between cell contractility and cancer invasiveness in bladder cancer (BLCA) remains understudied, despite the fact that BLCA is the 6th most prevalent cancer with high risk of recurrence. Here, we explore the relationship between the contractile phenotype of cancer cells and their invasiveness.

The invasive potential of five BLCA cell lines (RT4, SW-780, SW-1710, MGH-U3 and T24) was investigated using transwell and 3D spheroid invasion assays. Cell contractility was measured using traction force microscopy, 3D collagen gel compaction and quantitative polarized light microscopy of spheroids. Mechanotransduction was measured by YAP nuclear translocation and stiffness-mediated cell migration was investigated on stiffness-gradient substrates.

We found that the T24 and MGH-U3 cell lines were capable of invasion, while SW-780, RT4, and SW-1710 were non-invasive. The invasive cell lines exerted higher traction forces on their surrounding environment compared to their non-invasive counterparts. Furthermore, the invasive cell lines exhibited increased YAP nuclear translocation on stiffer substrate. When seeded on a stiffness-gradient substrate, invasive cell lines preferred high stiffnesses, whereas non-invasive cell lines migrated towards low-to-mid stiffness substrates.

Our findings uncover the relationship between the invasiveness of BLCA cell lines and their contractile phenotypes. We also show that a single assay is not sufficient to infer on a given aggressive phenotype. Furthermore, this study provides a comparative baseline for these cell lines to expand mechanobiology-related investigation in BLCA.

The online version contains supplementary material available at 10.1007/s12195-026-00897-y.