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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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Decreased expression of 37/67LR results in reduction of cell adhesion to LM-111 YIGSR related peptide.Controls and HIEC cells exhibiting reduced 37/67LR expression (siLR4-20) were used for 1h adhesion assays on low-fouling carboxymethyl-dextran (CMD) layers bearing the tripeptide Arg–Gly–Asp (RGD) or CDPGYIGSR, a laminin nonapeptide (YIGSR). The CMD surface prevents cell adhesion. HIEC cell adhesion to RGD was maximal and comparable for control and siLR-treated cells whereas a statistically significant reduction of cell adhesion on YIGSR was observed in partially 37/67LR-depleted cells (mean ± SEM, n=3, ** p < 0.01).
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pone-0074337-g007: Decreased expression of 37/67LR results in reduction of cell adhesion to LM-111 YIGSR related peptide.Controls and HIEC cells exhibiting reduced 37/67LR expression (siLR4-20) were used for 1h adhesion assays on low-fouling carboxymethyl-dextran (CMD) layers bearing the tripeptide Arg–Gly–Asp (RGD) or CDPGYIGSR, a laminin nonapeptide (YIGSR). The CMD surface prevents cell adhesion. HIEC cell adhesion to RGD was maximal and comparable for control and siLR-treated cells whereas a statistically significant reduction of cell adhesion on YIGSR was observed in partially 37/67LR-depleted cells (mean ± SEM, n=3, ** p < 0.01).

Mentions: Previous studies have identified that 37/67LR binds to the short arm of the laminin β chain via the YIGSR peptide sequence [11]. This characteristic was used to evaluate whether 37/67LR functions as a laminin-111 receptor in normal intestinal epithelial cells. Cell adhesion assays were performed on low-fouling surfaces on which peptide immobilization supports specific cell-biomaterial interaction while preventing non-specific protein adsorption allowing the discrimination of a specific from a non-specific cell response [72]. As performed previously [73], non-specific adhesion was determined on CMD surfaces which prevent cell adhesion, 37/67LR-mediated adhesion was evaluated on CDPGYIGSR (YIGSR) while maximal adhesion was determined on RGD peptide. Previous studies have shown that HIEC cells attach strongly to RGD peptides [71]. The results showed that reduced 37/67LR expression had no significant effect on binding to the RGD peptide but induced a statistically significant ~50% reduction of adhesion to YIGSR (Figure 7) confirming that 37/67LR is functional for LM-111 adhesion in intestinal crypt cells.


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)

Decreased expression of 37/67LR results in reduction of cell adhesion to LM-111 YIGSR related peptide.Controls and HIEC cells exhibiting reduced 37/67LR expression (siLR4-20) were used for 1h adhesion assays on low-fouling carboxymethyl-dextran (CMD) layers bearing the tripeptide Arg–Gly–Asp (RGD) or CDPGYIGSR, a laminin nonapeptide (YIGSR). The CMD surface prevents cell adhesion. HIEC cell adhesion to RGD was maximal and comparable for control and siLR-treated cells whereas a statistically significant reduction of cell adhesion on YIGSR was observed in partially 37/67LR-depleted cells (mean ± SEM, n=3, ** p < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0074337-g007: Decreased expression of 37/67LR results in reduction of cell adhesion to LM-111 YIGSR related peptide.Controls and HIEC cells exhibiting reduced 37/67LR expression (siLR4-20) were used for 1h adhesion assays on low-fouling carboxymethyl-dextran (CMD) layers bearing the tripeptide Arg–Gly–Asp (RGD) or CDPGYIGSR, a laminin nonapeptide (YIGSR). The CMD surface prevents cell adhesion. HIEC cell adhesion to RGD was maximal and comparable for control and siLR-treated cells whereas a statistically significant reduction of cell adhesion on YIGSR was observed in partially 37/67LR-depleted cells (mean ± SEM, n=3, ** p < 0.01).
Mentions: Previous studies have identified that 37/67LR binds to the short arm of the laminin β chain via the YIGSR peptide sequence [11]. This characteristic was used to evaluate whether 37/67LR functions as a laminin-111 receptor in normal intestinal epithelial cells. Cell adhesion assays were performed on low-fouling surfaces on which peptide immobilization supports specific cell-biomaterial interaction while preventing non-specific protein adsorption allowing the discrimination of a specific from a non-specific cell response [72]. As performed previously [73], non-specific adhesion was determined on CMD surfaces which prevent cell adhesion, 37/67LR-mediated adhesion was evaluated on CDPGYIGSR (YIGSR) while maximal adhesion was determined on RGD peptide. Previous studies have shown that HIEC cells attach strongly to RGD peptides [71]. The results showed that reduced 37/67LR expression had no significant effect on binding to the RGD peptide but induced a statistically significant ~50% reduction of adhesion to YIGSR (Figure 7) confirming that 37/67LR is functional for LM-111 adhesion in intestinal crypt cells.

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