Netrin-1 Induces MMP-12-Dependent E-Cadherin Degradation Via the Distinct Activation of PKCα and FAK/Fyn in Promoting Mesenchymal Stem Cell Motility

Abstract

Netrin-1 (Ntn-1) is a potent inducer of neuronal cell migration; however, its molecular mechanism that guides the migratory behavior of stem cells has not been characterized. In this study, we investigate the role of Ntn-1 in promoting the motility of human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) and its related signaling pathways. Ntn-1 (50 ng/mL) significantly increased motility of UCB-MSCs, which was inhibited by blocking antibodies for deleted in colorectal cancer (DCC) and integrin (IN) a6b4. Ntn-1 in DCC stimulated protein kinase Ca (PKCa) activation, but not PKCe, PKCy, and PKCz, while Ntn-1 in INa6b4 induced the phosphorylation of focal adhesion kinase (FAK) and Fyn. Notably, Ntn-1 induced phosphorylation of extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases ( JNK), and nuclear factor kappa-B (NF-kB), but they were concurrently downregulated by blocking the activities of PKCa, FAK, and Fyn. Ntn-1 uniquely increased the MMP-12 expression of all the matrix metalloproteinase (MMP) isoforms present in UCB-MSCs, though this was significantly blocked by an NF-kB inhibitor. Finally, Ntn-1 induced the MMP-12-dependent degradation of E-cadherin (E-cad), while Ntn-1 abrogated the interaction between E-cad and p120-catenin. In addition, Ntn-1 has the ability to stimulate cytoskeletal reorganization-related proteins, such as Cdc42, Rac1, Profilin-1, Cofilin-1, a-Actinin-4, and filamentous actin (F-actin) in UCB-MSCs. These results demonstrate that Ntn-1 induces MMP-12-dependent E-cad degradation via the distinct activation of PKCa and FAK/Fyn, which is necessary to govern the activation of ERK, JNK, and NF-kB in promoting motility of UCBMSCs.

Oleic acid enhances the motility of umbilical cord blood derived mesenchymal stem cells through EphB2-dependent F-actin formation

“…Because PKCα is a conventional PKC that requires both Ca 2+ and DAG for activation [46], Ca 2+ binding to the C2 domain of matured PKCα initiates translocation to the membrane. And several studies have reported that PKCα activation plays a crucial role in the migration of cord-bloodderived stem cells [47,48]. Related to this, OA-induced PKCα phosphorylation was inhibited by a GPR40 antagonist, GW1100, and controlled by Gα q , but not by Gα i or Gα 12 .…”

High Glucose-Induced Reactive Oxygen Species Stimulates Human Mesenchymal Stem Cell Migration Through Snail and EZH2-Dependent E-Cadherin Repression

“…Thus, migration is considered to be an important factor of MSC-induced wound healing. E-cadherin degradation by netrin pretreatment enhances migratory ability of hUCB-MSCs [50]. The reduction of E-cadherin expression during cell migration can be controlled by epigenetic transcriptional regulation and post-transcriptional modification of transcription for the CDH1 gene [51].…”

miR-142 acts as a tumor suppressor in osteosarcoma cell lines by targeting Rac1

“…E-cadherin is frequently downregulated during carcinoma metastasis (26). Loss of E-cadherin facilitates the initial invasive behavior of cancer (27). MMPs promote cell invasion and migration in several types of cancers, such as osteosarcoma, prostate, lung, colon and pancreas cancer (28).…”