Anin vitrocorrelation of mechanical forces and metastatic capacity

Abstract

Mechanical forces have a major influence on cell migration and are predicted to significantly impact cancer metastasis, yet this idea is currently poorly defined. In this study we have asked if changes in traction stress and migratory properties correlate with the metastatic progression of tumor cells. For this purpose, four murine breast cancer cell lines derived from the same primary tumor, but possessing increasing metastatic capacity, were tested for adhesion strength, traction stress, focal adhesion organization and for differential migration rates in two-dimensional and three-dimensional environments. Using traction force microscopy (TFM), we were surprised to find an inverse relationship between traction stress and metastatic capacity, such that force production decreased as the metastatic capacity increased. Consistent with this observation, adhesion strength exhibited an identical profile to the traction data. A count of adhesions indicated a general reduction in the number as metastatic capacity increased but no difference in the maturation as determined by the ratio of nascent to mature adhesions. These changes correlated well with a reduction in active beta-1 integrin with increasing metastatic ability. Finally, in two dimensions, wound healing, migration and persistence were relatively low in the entire panel, maintaining a downward trend with increasing metastatic capacity. Why metastatic cells would migrate so poorly prompted us to ask if the loss of adhesive parameters in the most metastatic cells indicated a switch to a less adhesive mode of migration that would only be detected in a three-dimensional environment. Indeed, in three-dimensional migration assays, the most metastatic cells now showed the greatest linear speed. We conclude that traction stress, adhesion strength and rate of migration do indeed change as tumor cells progress in metastatic capacity and do so in a dimension-sensitive manner.

Cell membrane topology analysis by RICM enables marker-free adhesion strength quantification

“…It is known that cell-substrate adhesion strength decreases as metastatic capacity increases [38]. This finding suggests that as cells progress in their metastatic capacity they produce fewer focal adhesions (FAs), perhaps resulting in a less adhesion-dependent mode of migration.…”

“…The results of these investigations are in good agreement with traction force studies showing that traction stress in two-dimensional cultures and cell-substrate adhesion strength are decreased as metastatic capacity increases. In addition, a reduction in the overall number of focal adhesions and the level of active beta-1 integrin are indicators of an increased metastatic capacity of the cell [38]. Because there is a linear relationship between the internal forces exerted by a focal adhesion and its lateral size [39], the adhesion strength can potentially be used to estimate the invasiveness of tumor cells.…”

Fluorescence-Based Force/Tension Sensors: A Novel Tool to Visualize Mechanical Forces in Structural Proteins in Live Cells

“…Cancer cells respond to mechanical stresses during tumor inception, transformation, proliferation, and metastasis (42,93). Metastasis is the major cause of cancer death, accounting for 90% of fatalities.…”

Polyacrylamide Gels for Invadopodia and Traction Force Assays on Cancer Cells

“…While actomyosin contractility appears to be important in this process, current studies conflict as to whether metastatic capacity is correlated to increased or decreased contractile forces 6,[18][19][20] . Furthermore, it remains unknown whether these forces directly mediate invadopodia activity 21 .…”