Assessing metastatic potential of breast cancer cells based on EGFR dynamics

Yen-Liang Liu,Andrew K Dunn,Yu-An Kuo,Chao-Kai Chou,Mirae Kim,Phyllis Ang,Katherine Blocher,Rohan Vasisht,Evan P Perillo,Mien-Chie Hung,Hsin-Chih Yeh,Cong Liu,Thomas E Yankeelov,Hannah Horng,Yu-An Chen,Yuan-I Chen,Duc Trung Nguyen

doi:10.1038/s41598-018-37625-0

Abstract

Derailed transmembrane receptor trafficking could be a hallmark of tumorigenesis and increased tumor invasiveness, but receptor dynamics have not been used to differentiate metastatic cancer cells from less invasive ones. Using single-particle tracking techniques, we developed a phenotyping asssay named Transmembrane Receptor Dynamics (TReD), studied the dynamics of epidermal growth factor receptor (EGFR) in seven breast epithelial cell lines and developed a phenotyping assay named Transmembrane Receptor Dynamics (TReD). Here we show a clear evidence that increased EGFR diffusivity and enlarged EGFR confinement size in the plasma membrane (PM) are correlated with the enhanced metastatic potential in these cell lines. By comparing the TReD results with the gene expression profiles, we found a clear negative correlation between the EGFR diffusivities and the breast cancer luminal differentiation scores (r = −0.75). Upon the induction of epithelial-mesenchymal transition (EMT), EGFR diffusivity significantly increased for the non-tumorigenic MCF10A (99%) and the non-invasive MCF7 (56%) cells, but not for the highly metastatic MDA-MB-231 cell. We believe that the reorganization of actin filaments during EMT modified the PM structures, causing the receptor dynamics to change. TReD can thus serve as a new biophysical marker to probe the metastatic potential of cancer cells and even to monitor the transition of metastasis.

Highlights

Receptor tyrosine kinases (RTKs) control many cell decision-making functions such as proliferation, survival, and movement
We demonstrate that epidermal growth factor receptor (EGFR) dynamics, as an example of Transmembrane Receptor Dynamics (TReD), can be used to differentiate breast cell lines with distinct metastatic potential and monitor the epithelial-mesenchymal transition in the benign cell line
We believe EGFR dynamics are coupled to the signaling networks through the local actin environment of the cancer cells, and changes in cancer cell behaviors, such as epithelial-mesenchymal transition, can alter the EGFR dynamics (Fig. 1A)

Summary

Introduction

Receptor tyrosine kinases (RTKs) control many cell decision-making functions such as proliferation, survival, and movement. To provide a multifaceted description of cancer cells, researchers have recently begun to explore physical properties of cancer cells (e.g., morphology[4], viscoelasticity[5], shear rheology[6], and motility7), with a hope to find an alternative way to quickly and precisely identify highly invasive cancer subtypes[8,9] These physical science approaches have revealed dramatic differences in mechanics, migration, and adhesion between MCF10A (non-tumorigenic) and MDA-MB-231 (highly invasive) breast cell lines[8]. There is no physical interrogation technique that overcomes all of the above issues To address this challenge, we have developed a new biophysical phenotyping method termed Transmembrane Receptor Dynamics (TReD), and showed that changes of TReD can be a signature of increased invasiveness. While our results agree well with the previous reports, our TReD assay is substantially easier than the current methods

Methods

Results

Conclusion