Suppression of cell migration by phospholipase C-related catalytically inactive protein-dependent modulation of PI3K signalling

Abstract

The metabolic processes of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] into PI(3,4,5)P3 and the subsequent PI(3,4,5)P3 signalling are involved in cell migration. Dysfunctions in the control of this pathway can cause human cancer cell migration and metastatic growth. Here we investigated whether phospholipase C-related catalytically inactive protein (PRIP), a PI(4,5)P2-binding protein, regulates cancer cell migration. PRIP overexpression in MCF-7 and BT-549 human breast cancer cells inhibited cell migration in vitro and metastasis development in vivo. Overexpression of the PRIP pleckstrin homology domain, a PI(4,5)P2 binding motif, in MCF-7 cells caused significant suppression of cell migration. Consistent with these results, in comparison with wild-type cells, Prip-deficient mouse embryonic fibroblasts exhibited increased cell migration, and this was significantly attenuated upon transfection with a siRNA targeting p110α, a catalytic subunit of class I phosphoinositide 3-kinases (PI3Ks). PI(3,4,5)P3 production was decreased in Prip-overexpressing MCF-7 and BT-549 cells. PI3K binding to PI(4,5)P2 was significantly inhibited by recombinant PRIP in vitro, and thus the activity of PI3K was downregulated. Collectively, PRIP regulates the production of PI(3,4,5)P3 from PI(4,5)P2 by PI3K, and the suppressor activity of PRIP in PI(4,5)P2 metabolism regulates the tumour migration, suggesting PRIP as a promising target for protection against metastatic progression.

Phospholipase C-related catalytically inactive protein regulates cytokinesis by protecting phosphatidylinositol 4,5-bisphosphate from metabolism in the cleavage furrow

“…In addition, PI(4,5)P 2 regulates the accumulation of RhoA at the cleavage furrow in HeLa cells 3 . We previously reported a protective effect of PRIP on PI(4,5)P 2 metabolism in migrating cells 27 . Therefore, we predicted that PI(4,5)P 2 accumulation at the cleavage furrow would be regulated by PRIP.…”

“…To investigate the effects of PRIP in OCRL1-expressed cells on cytokinesis progression, we performed live-cell imaging. We used MCF-7 cells stably expressing Prip1 , Prip1(R134Q) , or EGFP vector 27 in which Halo vector or Halo- OCRL1 was transiently transfected. Approximately 80% of OCRL1 -transfected EGFP vector-expressing MCF-7 cells showed abnormal cytokinesis (Fig.…”

“…PI(4,5)P 2 interacts with Rho, regulates Rho GTPase activity at the cleavage furrow, and facilitates the recruitment of contractile ring constituents to the cleavage furrow as a scaffold lipid 3,9 . We previously reported that PRIP negatively regulates PI(4,5)P 2 metabolism in migrating cells 27 . In this study, a deficiency of PRIP2 in HeLa cells was found to cause cytokinesis defects and delay by suppressing the accumulation of PI(4,5)P 2 and RhoA at the cleavage furrow.…”

“…We recently reported that PRIP negatively regulates the conversion of PI(4,5)P 2 into phosphatidylinositol 3,4,5-trisphosphate by phosphoinositide 3-kinase (PI3K), a process that suppresses the migration of cancer cells 27 . However, little is known regarding the involvement of PRIP in PI(4,5)P 2 -dependent cytokinetic events.…”

Leader cell PLCγ1 activation during keratinocyte collective migration is induced by EGFR localization and clustering

“…Thus, PLCγ1 may serve as a target for improving wound re‐epithelialization, and understanding the factors that lead to its activation is a crucial step toward developing therapies or wound dressings capable of modulating this signal. Additionally, PLCγ1 is recognized to impact motility in a wide range of cell types, therefore, elucidation of the factors that cause edge‐specific activation of PLCγ1 may translate to other cell types and tissues.…”