Protein kinases C-γ and -δ are involved in insulin-like growth factor I–induced migration of colonic epithelial cells

André, Frédéric; Rigot, Véronique; Remacle‐Bonnet, Maryse; Luis, José; Pommier, Gilbert; Marvaldi, Jacques

doi:10.1016/s0016-5085(99)70230-1

Cited by 71 publications

(74 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Wound Healing Assay-Population-based movement of cells was measured by a wound healing assay as described previously (29). In this assay, cells are grown to confluency on plastic Petri dishes, then induced to re-populate a "wound" created by stripping a sharply defined channel using a sterile razor blade.…”

Section: Methodsmentioning

confidence: 99%

Two Novel Functions of Hyaluronidase-2 (Hyal2) Are Formation of the Glycocalyx and Control of CD44-ERM Interactions

Duterme

Mertens-Strijthagen

Tammi

et al. 2009

Journal of Biological Chemistry

View full text Add to dashboard Cite

It has long been predicted that the members of the hyaluronidase enzyme family have important non-enzymatic functions. However, their nature remains a mystery. The metabolism of hyaluronan (HA), their major enzymatic substrate, is also enigmatic. To examine the function of Hyal2, a glycosylphosphatidylinositol-anchored hyaluronidase with intrinsically weak enzymatic activity, we have compared stably transfected rat fibroblastic BB16 cell lines with various levels of expression of Hyal2. These cell lines continue to express exclusively the standard form (CD44s) of the main HA receptor, CD44. Hyal2, CD44, and one of its main intracellular partners, ezrin-radixin-moesin (ERM), were found to co-immunoprecipitate. Functionally, Hyal2 overexpression was linked to loss of the glycocalyx, the HA-rich pericellular coat. This effect could be mimicked by exposure of BB16 cells either to Streptomyces hyaluronidase, to HA synthesis inhibitors, or to HA oligosaccharides. This led to shedding of CD44, separation of CD44 from ERM, reduction in baseline level of ERM activation, and markedly decreased cell motility (50% reduction in a wound healing assay). The effects of Hyal2 on the pericellular coat and on CD44-ERM interactions were inhibited by treatment with the Na ؉ /H ؉ exchanger-1 inhibitor ethyl-N-isopropylamiloride. We surmise that Hyal2, through direct interactions with CD44 and possibly some pericellular hyaluronidase activity requiring acidic foci, suppresses the formation or the stability of the glycocalyx, modulates ERMrelated cytoskeletal interactions, and diminishes cell motility. These effects may be relevant to the purported in vivo tumorsuppressive activity of Hyal2.

show abstract

Section: Methodsmentioning

confidence: 99%

Two Novel Functions of Hyaluronidase-2 (Hyal2) Are Formation of the Glycocalyx and Control of CD44-ERM Interactions

Duterme

Mertens-Strijthagen

Tammi

et al. 2009

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Each coverslip was washed twice and incubated for 15 min further in calcium buffer at room temperature and then transferred to a quartz cuvette containing 2 ml of the same buffer at 37°C. After 2 min of equilibration, fluorescence was measured via a Hitachi F-2000 fluorospectrophotometer with dual-excitation wavelengths of 340 nm ( 1) and 380 nm ( 2) and an emission wavelength of 510 nM while the incubation medium was continually stirred at 37°C. [Ca 2ϩ ]i was determined from the formula…”

Section: Methodsmentioning

confidence: 99%

“…INTESTINAL EPITHELIAL CELL FUNCTIONS, including growth, motility, differentiation, and transport, are regulated by a combination of environmental factors that include peptide signals, bioactive lipids, adhesion to the extracellular matrix, and cell wounding (2,7,13,33,47,55). The nontransformed IEC-18 and IEC-6 intestinal epithelial cell lines, derived from rat small intestinal crypt epithelium (40), have been widely used as a model of epithelial cell proliferation, migration, and differentiation (7,9,33,55).…”

mentioning

confidence: 99%

ANG II and LPA induce Pyk2 tyrosine phosphorylation in intestinal epithelial cells: role of Ca²⁺, PKC, and Rho kinase

Wu¹,

Chiu

Rozengurt

2002

American Journal of Physiology-Cell Physiology

View full text Add to dashboard Cite

2001.-The G protein-coupled receptor agonists angiotensin II (ANG II) and lysophosphatidic acid (LPA) rapidly induce tyrosine phosphorylation of the cytosolic proline-rich tyrosine kinase 2 (Pyk2) in IEC-18 intestinal epithelial cells. The combined Pyk2 tyrosine phosphorylation induced by phorbol 12,13-dibutyrate, a direct agonist of protein kinase C (PKC), and ionomycin, a Ca 2ϩ ionophore, was equal to that induced by ANG II. Inhibition of either PKC or Ca 2ϩ signaling attenuated the effect of ANG II and LPA, although simultaneous inhibition of both pathways failed to completely abolish Pyk2 tyrosine phosphorylation. Cytochalasin D, which disrupts stress fibers, strongly inhibited the response of Pyk2 to ANG II or LPA. The distinct Rho-associated kinase (ROK) inhibitors HA-1077 and Y-27632, as well as the Rho inhibitor Clostridium botulinum C3 exoenzyme, also significantly attenuated ANG II-and LPA-stimulated Pyk2 tyrosine phosphorylation. Simultaneous inhibition of PKC, Ca 2ϩ , and either actin assembly or ROK completely abolished the Pyk2 response. Together, these results show that ANG II and LPA rapidly induce Pyk2 tyrosine phosphorylation in intestinal epithelial cells via separate Ca 2ϩ -, PKC-, and Rho-mediated pathways.

show abstract

“…Migration of cells used in this study requires activation of PKC, either by PMA or by growth factors. 30,33 Hepatocyte growth factor (HGF) is produced by various cell types, including tumor-bordering myofibroblasts, and functions locally as a cell migration inductor. As illustrated in Figure 3A, HGF promoted cell migration to vitronectin more efficiently than PMA.…”

Section: Proprotein Convertases Inhibition Increases Cell Migrationmentioning

confidence: 99%

Inhibition of Proprotein Convertases Enhances Cell Migration and Metastases Development of Human Colon Carcinoma Cells in a Rat Model

Nejjari

Berthet

Rigot

et al. 2004

The American Journal of Pathology

Self Cite

View full text Add to dashboard Cite

Although proprotein convertases are involved in tumor development, nothing is known about their role in metastatic dissemination. To investigate the involvement of convertase inhibition, we used human colon carcinoma cells overexpressing ␣1-antitrypsin Portland (␣1-PDX, PDX39P cells), a potent convertase inhibitor. We previously reported that these cells bear uncleaved integrin ␣ subunits and display an altered attachment to vitronectin that is correlated with defects in the intracellular signaling pathways activated by ␣v␤5 integrin ligation. In this study, we demonstrate that the inhibition of proprotein convertase activity either by overexpression of ␣1-PDX or with the synthetic inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (dec-RVKR-cmk) led to a significant increase in cell migration supported by the ␣v␤5 integrin. A collagen gel invasion assay showed that PDX39P cells also displayed an invasive ability, contrary to control cells. Moreover, when injected to immunosuppressed newborn rats, PDX39P cells were highly invasive, as they induce 10 times more metastases than mock-transfected cells. In addition, the aggressiveness of PDX39P cells can be greatly reduced by a function-blocking monoclonal antibody (mAb) against the ␣v subunit. It thus seems that inhibition of proprotein convertases enhances the in vivo invasiveness of colon tumor cells likely due to an increase in cell migration mediated by ␣v integrins. The acquisition of cell motility and the capacity to invade basement matrix membranes and adjacent tissues play a central role in the complex multi-step process of metastasis. Cell migration is the result of dynamic interactions between the cell, the extracellular matrix (ECM), and the cytoskeleton. Integrins connect the ECM proteins outside to the actin cytoskeleton within the cell, allowing the traction required for cell migration. 1,2 Integrins also play a crucial role in signal transduction events that control anchorage-independent growth and survival of tumor cells (for reviews, see references [3][4][5] ). Therefore, these adhesion molecules play a pivotal role in tumor cell invasion and metastasis. [5][6][7] Integrins are heterodimeric proteins composed by the non-covalent association of ␣ and ␤ subunits. Some integrin ␣ chains undergo a post-translational cleavage in their extracellular domain by furin and PC5A, two proprotein convertases (PCs) of the subtilisin/kexin family. 8,9 The role of this cleavage in integrin function remains unclear. Arguably, this post-translational processing of the ␣ chain is not required for ligand binding, 10 -12 but is essential for the signaling function of ␣6␤1 and ␣v␤5 integrins. 10,13 Moreover, recent data suggest that the ␣6 subunit cleavage might be linked to the differentiation state of lens cells. 14 PCs are responsible for the biological activation of a variety of precursor proteins, including pro-hormones and their receptors and adhesion molecules. They are ubiquitously expressed and are involved in many physiological and pathological processes. ...

show abstract

Protein kinases C-γ and -δ are involved in insulin-like growth factor I–induced migration of colonic epithelial cells

Cited by 71 publications

References 47 publications

Two Novel Functions of Hyaluronidase-2 (Hyal2) Are Formation of the Glycocalyx and Control of CD44-ERM Interactions

Two Novel Functions of Hyaluronidase-2 (Hyal2) Are Formation of the Glycocalyx and Control of CD44-ERM Interactions

ANG II and LPA induce Pyk2 tyrosine phosphorylation in intestinal epithelial cells: role of Ca²⁺, PKC, and Rho kinase

Inhibition of Proprotein Convertases Enhances Cell Migration and Metastases Development of Human Colon Carcinoma Cells in a Rat Model

Contact Info

Product

Resources

About

Protein kinases C-γ and -δ are involved in insulin-like growth factor I–induced migration of colonic epithelial cells

Cited by 71 publications

References 47 publications

Two Novel Functions of Hyaluronidase-2 (Hyal2) Are Formation of the Glycocalyx and Control of CD44-ERM Interactions

Two Novel Functions of Hyaluronidase-2 (Hyal2) Are Formation of the Glycocalyx and Control of CD44-ERM Interactions

ANG II and LPA induce Pyk2 tyrosine phosphorylation in intestinal epithelial cells: role of Ca2+, PKC, and Rho kinase

Inhibition of Proprotein Convertases Enhances Cell Migration and Metastases Development of Human Colon Carcinoma Cells in a Rat Model

Contact Info

Product

Resources

About

ANG II and LPA induce Pyk2 tyrosine phosphorylation in intestinal epithelial cells: role of Ca²⁺, PKC, and Rho kinase