open access

Vol 54, No 2 (2016)
Original paper
Submitted: 2016-05-14
Accepted: 2016-06-27
Published online: 2016-07-13
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Myenteric plexuses atrophy in the vicinity of colorectal cancer tissue is not caused by apoptosis or necrosis

Anna Kozlowska, Przemyslaw Kwiatkowski, Agnieszka Oponowicz, Mariusz Majewski, Zbigniew Kmiec, Janusz Godlewski
DOI: 10.5603/FHC.a2016.0012
·
Pubmed: 27439439
·
Folia Histochem Cytobiol 2016;54(2):99-107.

open access

Vol 54, No 2 (2016)
ORIGINAL PAPERS
Submitted: 2016-05-14
Accepted: 2016-06-27
Published online: 2016-07-13

Abstract

Introduction. The previously performed studies showed that the presence of colorectal cancer (CRC) tumor is associated with the atrophy of myenteric plexuses in the vicinity of cancer invasion; however, the possible mechanisms of this phenomenon are not known. The aim of the present study was to determine whether the atrophic changes of the enteric nervous system (ENS) within an intestine wall of the CRC patients were caused by apoptosis or necrosis and whether they were associated with changes in the number of galanin-immunore­active (GAL-Ir) neurons.

Material and methods. Samples of the large intestine wall located close to the CRC invasion and control, distally-located part of the colon, were collected from 9 CRC patients. The size of ENS plexuses and the number of neurons were compared. Triple immunofluorescent staining was used to visualize the co-expression of caspase 3 (CASP3) or caspase 8 (CASP8) with GAL and protein gene-product 9.5 (PGP 9.5, panneuronal marker) in the submucosal and myenteric ENS plexuses. The cells expressing myeloperoxidase (MPO, marker of neutrophils) and CD68 (marker of macrophages) were detected by immunohistochemistry around/in myenteric plexuses (MPs) and in the muscularis externa of the colon wall in the vicinity of tumor invasion.

Results. Myenteric plexuses in the vicinity of the CRC tissue were significantly smaller and had lower number of neurons per plexus than distantly located plexuses. The number of CASP8- and CASP3-Ir neurons in the ENS plexuses was similar in the colon wall both close to and distally from tumor invasion. The number of CASP8-Ir neurons within MPs located close to CRC invasion was higher than of CASP3-Ir neurons. The percentage of neurons co-expressing CASP8 and GAL in myenteric plexuses close and distantly from tumor was three-fold lower than of those co-expressing CASP3 and GAL. The mean number of neutrophils and macrophages inside and around myenteric plexuses located close to tumor invasion was higher or similar, respectively, as compared with adjacent muscularis externa.

Conclusions. The atrophy of myenteric plexuses in the vicinity of CRC invasion is not caused by apoptosis or necrosis. The differences in the proportions of neurons expressing galanin and the studied caspases suggest as yet unknown role of this neuropeptide in the mechanisms of neuron’s atrophy in MPs located close to CRC tumor.

Abstract

Introduction. The previously performed studies showed that the presence of colorectal cancer (CRC) tumor is associated with the atrophy of myenteric plexuses in the vicinity of cancer invasion; however, the possible mechanisms of this phenomenon are not known. The aim of the present study was to determine whether the atrophic changes of the enteric nervous system (ENS) within an intestine wall of the CRC patients were caused by apoptosis or necrosis and whether they were associated with changes in the number of galanin-immunore­active (GAL-Ir) neurons.

Material and methods. Samples of the large intestine wall located close to the CRC invasion and control, distally-located part of the colon, were collected from 9 CRC patients. The size of ENS plexuses and the number of neurons were compared. Triple immunofluorescent staining was used to visualize the co-expression of caspase 3 (CASP3) or caspase 8 (CASP8) with GAL and protein gene-product 9.5 (PGP 9.5, panneuronal marker) in the submucosal and myenteric ENS plexuses. The cells expressing myeloperoxidase (MPO, marker of neutrophils) and CD68 (marker of macrophages) were detected by immunohistochemistry around/in myenteric plexuses (MPs) and in the muscularis externa of the colon wall in the vicinity of tumor invasion.

Results. Myenteric plexuses in the vicinity of the CRC tissue were significantly smaller and had lower number of neurons per plexus than distantly located plexuses. The number of CASP8- and CASP3-Ir neurons in the ENS plexuses was similar in the colon wall both close to and distally from tumor invasion. The number of CASP8-Ir neurons within MPs located close to CRC invasion was higher than of CASP3-Ir neurons. The percentage of neurons co-expressing CASP8 and GAL in myenteric plexuses close and distantly from tumor was three-fold lower than of those co-expressing CASP3 and GAL. The mean number of neutrophils and macrophages inside and around myenteric plexuses located close to tumor invasion was higher or similar, respectively, as compared with adjacent muscularis externa.

Conclusions. The atrophy of myenteric plexuses in the vicinity of CRC invasion is not caused by apoptosis or necrosis. The differences in the proportions of neurons expressing galanin and the studied caspases suggest as yet unknown role of this neuropeptide in the mechanisms of neuron’s atrophy in MPs located close to CRC tumor.

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Keywords

colorectal cancer; enteric nervous system; plexus; caspase 3; caspase 8; galanin; CD68; myeloperoxidase; IHC

About this article
Title

Myenteric plexuses atrophy in the vicinity of colorectal cancer tissue is not caused by apoptosis or necrosis

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 54, No 2 (2016)

Article type

Original paper

Pages

99-107

Published online

2016-07-13

DOI

10.5603/FHC.a2016.0012

Pubmed

27439439

Bibliographic record

Folia Histochem Cytobiol 2016;54(2):99-107.

Keywords

colorectal cancer
enteric nervous system
plexus
caspase 3
caspase 8
galanin
CD68
myeloperoxidase
IHC

Authors

Anna Kozlowska
Przemyslaw Kwiatkowski
Agnieszka Oponowicz
Mariusz Majewski
Zbigniew Kmiec
Janusz Godlewski

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