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The role of Wnt/β-catenin signaling in enterocyte turnover during methotrexate-induced intestinal mucositis in a rat.

Sukhotnik I, Geyer T, Pollak Y, Mogilner JG, Coran AG, Berkowitz D - PLoS ONE (2014)

Bottom Line: The Wnt signaling pathway is critical for establishing and maintaining the proliferative compartment of the intestine.In the vitro experiment, treatment with MTX resulted in marked decrease in early cell proliferation rates following by a 17-fold increase in late cell proliferation rates compared to early proliferation.Treatment with MTX resulted in a significant increase in early and late apoptosis compared to Caco-2 untreated cells.

View Article: PubMed Central - PubMed

Affiliation: The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Laboratory of intestinal adaptation and recovery, Dept of Pediatric Surgery, Bnai Zion Medical Center, Haifa, Israel.

ABSTRACT

Background/aims: Intestinal mucositis is a common side-effect in patients who receive aggressive chemotherapy. The Wnt signaling pathway is critical for establishing and maintaining the proliferative compartment of the intestine. In the present study, we tested whether Wnt/β-catenin signaling is involved in methotrexate (MTX)-induced intestinal damage in a rat model.

Methods: Non-pretreated and pretreated with MTX Caco-2 cells were evaluated for cell proliferation and apoptosis using FACS analysis. Adult rats were divided into three experimental groups: Control rats; MTX-2 animals were treated with a single dose of MTX given IP and were sacrificed on day 2, and MTX-4 rats were treated with MTX similar to group B and were sacrificed on day 4. Intestinal mucosal damage, mucosal structural changes, enterocyte proliferation, and enterocyte apoptosis were measured at sacrifice. Real Time PCR and Western blot was used to determine the level of Wnt/β-catenin related genes and protein expression.

Results: In the vitro experiment, treatment with MTX resulted in marked decrease in early cell proliferation rates following by a 17-fold increase in late cell proliferation rates compared to early proliferation. Treatment with MTX resulted in a significant increase in early and late apoptosis compared to Caco-2 untreated cells. In the vivo experiment, MTX-2 and MTX-4 rats demonstrated intestinal mucosal hypoplasia. MTX-2 rats demonstrated a significant decrease in FRZ-2, Wnt 3A Wnt 5A, β-catenin, c-myc mRNA expression and a significant decrease in β-catenin and Akt protein levels compared to control animals. Four days following MTX administration, rats demonstrated a trend toward a restoration of Wnt/β-catenin signaling especially in ileum.

Conclusions: Wnt/β-catenin signaling is involved in enterocyte turnover during MTX-induced intestinal mucositis in a rat.

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Related in: MedlinePlus

Grade of intestinal mucosal injury and microscopic intestinal appearance after administration of MTX.Values are mean ± SEM. CONTR- control, MTX-methotrexate. * P<0.05 MTX-2 and MTX-4 vs CONTR rats, † P<0.05 MTX-4 vs MTX-2 rats.
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pone-0110675-g003: Grade of intestinal mucosal injury and microscopic intestinal appearance after administration of MTX.Values are mean ± SEM. CONTR- control, MTX-methotrexate. * P<0.05 MTX-2 and MTX-4 vs CONTR rats, † P<0.05 MTX-4 vs MTX-2 rats.

Mentions: The degree of intestinal damage increased following MTX administration, as indicated by severe villous atrophy, epithelial flattening, significant loss of crypt architecture, signs of crypt remodeling, severe villous epithelial atrophy, degeneration and shortening of the villus length, and polymorphonuclear leukocyte infiltration in the lamina propria. Two days following methotrexate administration, MTX-rats (Group B) demonstrated a significant increase in intestinal damage score in jejunum (6.2±0.9 vs 0.7±0.02, p<0.001) and distal ileum (6.2±0.2 vs 0.4±0.16, p<0.001) compared to control animals (Figure 3A). MTX-4 rats (Group C) demonstrated a more significant damage in the distal ileum (7.3±0.3 vs 6.2±0.3, p<0.05) compared to MTX-2 animals (Group B).


The role of Wnt/β-catenin signaling in enterocyte turnover during methotrexate-induced intestinal mucositis in a rat.

Sukhotnik I, Geyer T, Pollak Y, Mogilner JG, Coran AG, Berkowitz D - PLoS ONE (2014)

Grade of intestinal mucosal injury and microscopic intestinal appearance after administration of MTX.Values are mean ± SEM. CONTR- control, MTX-methotrexate. * P<0.05 MTX-2 and MTX-4 vs CONTR rats, † P<0.05 MTX-4 vs MTX-2 rats.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222808&req=5

pone-0110675-g003: Grade of intestinal mucosal injury and microscopic intestinal appearance after administration of MTX.Values are mean ± SEM. CONTR- control, MTX-methotrexate. * P<0.05 MTX-2 and MTX-4 vs CONTR rats, † P<0.05 MTX-4 vs MTX-2 rats.
Mentions: The degree of intestinal damage increased following MTX administration, as indicated by severe villous atrophy, epithelial flattening, significant loss of crypt architecture, signs of crypt remodeling, severe villous epithelial atrophy, degeneration and shortening of the villus length, and polymorphonuclear leukocyte infiltration in the lamina propria. Two days following methotrexate administration, MTX-rats (Group B) demonstrated a significant increase in intestinal damage score in jejunum (6.2±0.9 vs 0.7±0.02, p<0.001) and distal ileum (6.2±0.2 vs 0.4±0.16, p<0.001) compared to control animals (Figure 3A). MTX-4 rats (Group C) demonstrated a more significant damage in the distal ileum (7.3±0.3 vs 6.2±0.3, p<0.05) compared to MTX-2 animals (Group B).

Bottom Line: The Wnt signaling pathway is critical for establishing and maintaining the proliferative compartment of the intestine.In the vitro experiment, treatment with MTX resulted in marked decrease in early cell proliferation rates following by a 17-fold increase in late cell proliferation rates compared to early proliferation.Treatment with MTX resulted in a significant increase in early and late apoptosis compared to Caco-2 untreated cells.

View Article: PubMed Central - PubMed

Affiliation: The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Laboratory of intestinal adaptation and recovery, Dept of Pediatric Surgery, Bnai Zion Medical Center, Haifa, Israel.

ABSTRACT

Background/aims: Intestinal mucositis is a common side-effect in patients who receive aggressive chemotherapy. The Wnt signaling pathway is critical for establishing and maintaining the proliferative compartment of the intestine. In the present study, we tested whether Wnt/β-catenin signaling is involved in methotrexate (MTX)-induced intestinal damage in a rat model.

Methods: Non-pretreated and pretreated with MTX Caco-2 cells were evaluated for cell proliferation and apoptosis using FACS analysis. Adult rats were divided into three experimental groups: Control rats; MTX-2 animals were treated with a single dose of MTX given IP and were sacrificed on day 2, and MTX-4 rats were treated with MTX similar to group B and were sacrificed on day 4. Intestinal mucosal damage, mucosal structural changes, enterocyte proliferation, and enterocyte apoptosis were measured at sacrifice. Real Time PCR and Western blot was used to determine the level of Wnt/β-catenin related genes and protein expression.

Results: In the vitro experiment, treatment with MTX resulted in marked decrease in early cell proliferation rates following by a 17-fold increase in late cell proliferation rates compared to early proliferation. Treatment with MTX resulted in a significant increase in early and late apoptosis compared to Caco-2 untreated cells. In the vivo experiment, MTX-2 and MTX-4 rats demonstrated intestinal mucosal hypoplasia. MTX-2 rats demonstrated a significant decrease in FRZ-2, Wnt 3A Wnt 5A, β-catenin, c-myc mRNA expression and a significant decrease in β-catenin and Akt protein levels compared to control animals. Four days following MTX administration, rats demonstrated a trend toward a restoration of Wnt/β-catenin signaling especially in ileum.

Conclusions: Wnt/β-catenin signaling is involved in enterocyte turnover during MTX-induced intestinal mucositis in a rat.

Show MeSH
Related in: MedlinePlus