Screening for genes involved in collective cell migration

Background

The process of cell migration is important in diverse physiological and pathological processes such as development, immune surveillance and cancer metastasis. Cell migration at the single-cell level has been studied extensively over many decades. Collective cell migration is the second principal mode of cell movement, particularly prevalent during embryogenesis, where it drives the formation of complex tissues. This type of migration, also known as “invasion,” characterizes many invasive tumors. During collective cell migration, the cells remain physically and functionally connected, and the integrity of cell-cell junctions is preserved during movement, at least part of the time. Multicellular polarity and organization of the actin cytoskeleton generate traction and protrusion force for migration. In most modes of collective cell migration, moving cell groups modify the tissue along the migratory path (e.g., ECM modification and deposition of basement membrane).

Rationale

To better understand the process of collective cell migration and to find novel genes involved in this process, we’ve performed a siRNA screen on 4T1 mouse mammary tumor cell line - a highly metastatic breast cancer-derived cell line which displays collective migration.

Methods

Four Dharmacon Ltd. SMARTpool siRNA libraries (198 phosphatases, 719 kinases, 541 ubiquitination, and 112 glycoproteins) were used for targeting all assays (about 1,500 genes in total). Genes found to affect collective cell migration were assayed for their
involvement in other cancer cell types, and their mechanism of action was analyzed.

Contact person: Yair Elisha