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Laminin receptor 37/67LR regulates adhesion and proliferation of normal human intestinal epithelial cells.

Khalfaoui T, Groulx JF, Sabra G, GuezGuez A, Basora N, Vermette P, Beaulieu JF - PLoS ONE (2013)

Bottom Line: Immunolocalization studies revealed that 37/67LR was predominantly present in the undifferentiated/proliferative region of the human intestinal crypt in both the immature and adult intestine.Using a human intestinal epithelial crypt (HIEC) cell line as experimental model, we determined that 37/67LR was expressed in proliferative cells in both the cytoplasmic and membrane compartments.Small-interfering RNA-mediated reduction of 37/67LR expression led to HIEC cell-cycle reduction and loss of the ability to adhere to laminin-related peptides under conditions not altering ribosomal function.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Intestinal Physiopathology, Department of Anatomy and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada.

ABSTRACT
Interactions between the cell basal membrane domain and the basement membrane are involved in several cell functions including proliferation, migration and differentiation. Intestinal epithelial cells can interact with laminin, a major intestinal basement membrane glycoprotein, via several cell-surface laminin-binding proteins including integrin and non-integrin receptors. The 37/67kDa laminin receptor (37/67LR) is one of these but its role in normal epithelial cells is still unknown. The aim of this study was to characterise the expression pattern and determine the main function of 37/67LR in the normal human small intestinal epithelium. Immunolocalization studies revealed that 37/67LR was predominantly present in the undifferentiated/proliferative region of the human intestinal crypt in both the immature and adult intestine. Using a human intestinal epithelial crypt (HIEC) cell line as experimental model, we determined that 37/67LR was expressed in proliferative cells in both the cytoplasmic and membrane compartments. Small-interfering RNA-mediated reduction of 37/67LR expression led to HIEC cell-cycle reduction and loss of the ability to adhere to laminin-related peptides under conditions not altering ribosomal function. Taken together, these findings indicate that 37/67LR regulates proliferation and adhesion in normal intestinal epithelial cells independently of its known association with ribosomal function.

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Blocking 37/67LR reduces cell proliferation.HIEC cells were treated for 1h with neutralizing-blocking anti-37/67LR antibody (MLuC5) or anti-integrin β1 antibody (mAb13) as well as non-immune serum (NI) as negative control before plating. BrdU-positive cells were counted and expressed as percentage of total cells determined by DAPI staining (mean ± SEM, n=3, * p < 0.001; ** p < 0.0001).
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pone-0074337-g008: Blocking 37/67LR reduces cell proliferation.HIEC cells were treated for 1h with neutralizing-blocking anti-37/67LR antibody (MLuC5) or anti-integrin β1 antibody (mAb13) as well as non-immune serum (NI) as negative control before plating. BrdU-positive cells were counted and expressed as percentage of total cells determined by DAPI staining (mean ± SEM, n=3, * p < 0.001; ** p < 0.0001).

Mentions: Taking into consideration the presence of functional membrane 37/67LR receptor for laminin adhesion in HIEC cells, we further investigated the reduction of cell proliferation in 37/67LR depleted cells observed above (Figure 6) by conducting antibody blocking experiments on wild-type HIEC cells for a 24h period followed by BrdU incorporation. As shown in Figure 8, a significant reduction of BrdU incorporation was noted with the anti-37/67LR MLuC5 azide-free antibody. The effect appears to be specific since a non-immune serum (NI) had no effect. The blocking anti-β1 integrin mAb13 antibody [71] was used as a positive control.


Laminin receptor 37/67LR regulates adhesion and proliferation of normal human intestinal epithelial cells.

Khalfaoui T, Groulx JF, Sabra G, GuezGuez A, Basora N, Vermette P, Beaulieu JF - PLoS ONE (2013)

Blocking 37/67LR reduces cell proliferation.HIEC cells were treated for 1h with neutralizing-blocking anti-37/67LR antibody (MLuC5) or anti-integrin β1 antibody (mAb13) as well as non-immune serum (NI) as negative control before plating. BrdU-positive cells were counted and expressed as percentage of total cells determined by DAPI staining (mean ± SEM, n=3, * p < 0.001; ** p < 0.0001).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3750003&req=5

pone-0074337-g008: Blocking 37/67LR reduces cell proliferation.HIEC cells were treated for 1h with neutralizing-blocking anti-37/67LR antibody (MLuC5) or anti-integrin β1 antibody (mAb13) as well as non-immune serum (NI) as negative control before plating. BrdU-positive cells were counted and expressed as percentage of total cells determined by DAPI staining (mean ± SEM, n=3, * p < 0.001; ** p < 0.0001).
Mentions: Taking into consideration the presence of functional membrane 37/67LR receptor for laminin adhesion in HIEC cells, we further investigated the reduction of cell proliferation in 37/67LR depleted cells observed above (Figure 6) by conducting antibody blocking experiments on wild-type HIEC cells for a 24h period followed by BrdU incorporation. As shown in Figure 8, a significant reduction of BrdU incorporation was noted with the anti-37/67LR MLuC5 azide-free antibody. The effect appears to be specific since a non-immune serum (NI) had no effect. The blocking anti-β1 integrin mAb13 antibody [71] was used as a positive control.

Bottom Line: Immunolocalization studies revealed that 37/67LR was predominantly present in the undifferentiated/proliferative region of the human intestinal crypt in both the immature and adult intestine.Using a human intestinal epithelial crypt (HIEC) cell line as experimental model, we determined that 37/67LR was expressed in proliferative cells in both the cytoplasmic and membrane compartments.Small-interfering RNA-mediated reduction of 37/67LR expression led to HIEC cell-cycle reduction and loss of the ability to adhere to laminin-related peptides under conditions not altering ribosomal function.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Intestinal Physiopathology, Department of Anatomy and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada.

ABSTRACT
Interactions between the cell basal membrane domain and the basement membrane are involved in several cell functions including proliferation, migration and differentiation. Intestinal epithelial cells can interact with laminin, a major intestinal basement membrane glycoprotein, via several cell-surface laminin-binding proteins including integrin and non-integrin receptors. The 37/67kDa laminin receptor (37/67LR) is one of these but its role in normal epithelial cells is still unknown. The aim of this study was to characterise the expression pattern and determine the main function of 37/67LR in the normal human small intestinal epithelium. Immunolocalization studies revealed that 37/67LR was predominantly present in the undifferentiated/proliferative region of the human intestinal crypt in both the immature and adult intestine. Using a human intestinal epithelial crypt (HIEC) cell line as experimental model, we determined that 37/67LR was expressed in proliferative cells in both the cytoplasmic and membrane compartments. Small-interfering RNA-mediated reduction of 37/67LR expression led to HIEC cell-cycle reduction and loss of the ability to adhere to laminin-related peptides under conditions not altering ribosomal function. Taken together, these findings indicate that 37/67LR regulates proliferation and adhesion in normal intestinal epithelial cells independently of its known association with ribosomal function.

Show MeSH
Related in: MedlinePlus