Matrix metalloproteinase-26 (MMP-26) and Tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium.

Radovan Pilka

Matrix metalloproteinase-26 (MMP-26) and Tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium.

Radovan Pilka

Obstetrik och gynekologi, Lund

Forskningsoutput: Avhandling › Doktorsavhandling (sammanläggning)

Sammanfattning

Popular Abstract in Swedish
I avhandlingen studeras Matrix MetalloProteinase-26 (MMP-26) och Tissue Inhibitor of MetalloProteinases-4 (TIMP-4) i normalt, hyperplastiskt och malignt endometrium. Just detta protease/hämmarpar kan potentiellt vara av mycket stort intresse vid implantationsprocessen.

Trofoblastvävnad från såväl humant som murint embryo producerar proMMP-9. Aktivt MMP-9 (Gelatinase B) är ett proteolytiskt enzym med mycket bred specificitet. MMP-26 är en effektiv aktivator av proMMP-9, och TIMP-4 hämmar MMP-26 med mycket hög specificitet.

Vi har funnit att endometriets uttryck av de b?da generna för MMP-26 och TIMP-4 ökar under proliferations-fasen, är maximalt i tidig sekretionsfas, och faller därefter mycket snabbt i mid och sen sekretionsfas (I, III). Mönstret antyder att generna uppregleras av estradiol och nedregleras av progesterone. Detta antagande styrks av att vi identifierade s.k. estrogen response elements uppströms den kodande sekvensen i b?da generna (II, III). Med in situ hybridisering lokaliserade vi mRNA för MMP-26 till epitelet och TIMP-4 till stromat i normalt endometrium (I, III). MMP-26 protein återfanns som väntat i epitelet med immunhistokemi (II). Med samma teknik återfanns emellertid bara sparsamt TIMP-4 i stromat (V).

TIMP-4 lokaliserades i stället till epitelet, närmre bestämt granulaliknande strukturer apikalt i epitelcellerna. Fyndet antydde att TIMP-4 frisätts fr?n stromat, tages upp och ackumuleras i epitelcellerna, samt är där förberett för sekretion. I nästa steg undersöktes därför endometriesekretet (uterine fluid) med Western blot teknik. Vi fann TIMP-4, men inte MMP-26, i sekretet (V). Fyndet är intressant då mycket få protein i uterine fluid verkligen har visats häröra från stromat. Vidare fann vi med antikroppsteknik att MMP-26 upplagras i epitelcellerna i aktiverad form. MMP-26 frisätts inte spontant, men om så sker hämmas enzymet av TIMP-4 i s?väl stroma som uterine fluid.

Undersökning av endometrieprover fr?n patienter som genomg?r IVF behandling med substitution av b?de estradiol och progesterone, visar att östrogenkänsliga gener, som t.ex. MMP-26 och TIMP-4, överstimuleras först av östrogenet och därefter av progesterone.

Resultaten för både MMP-26 och TIMP-4 vid hyperplasi visar att både lokalisation och kvantitering är jämförbart med proliferationsfas. Detta passar med genernas östrogenreglering samt det faktum att hyperplasi är resultatet av överstimulering med östrogen.

Endometriecancer förlorar gradvis uttrycket av båda generna med tilltagande förlust av differentiering. Detta stämmer ocks? med en allmänt avtagande effekt av extern reglering då cancerceller förändras i riktning mot lägre differentierade, mer maligna tumörer.

Originalspråk	engelska
Kvalifikation	Doktor
Tilldelande institution	Obstetrik och gynekologi, Lund
Handledare	Casslén, Bertil, handledare
Tilldelningsdatum	2006 apr. 28
Förlag	Publishing Centre, Palacky University, Olomouc, Czech Republic
ISBN (tryckt)	91-85481-73-4
Status	Published - 2006

Bibliografisk information

Defence details

Date: 2006-04-28
Time: 13:00
Place: Dept. of Obstetrics and Gynecology, Lecture Hall

External reviewer(s)

Name: Marbaix, Etienne
Title: Professor
Affiliation: Department of Pathology, University of Louvain Medical School, Brussels, Belgium

---

<div class="article_info">R Pilka, C Whatling, H Domanski, S Hansson, P Eriksson and B Casslen. <span class="article_issue_date">2003</span>. <span class="article_title">Epithelial expression of matrix metalloproteinase-26 (MMP-26) is elevated at midcycle in the human endometrium.</span> <span class="journal_series_title">Mol Human Reprod,</span>, <span class="journal_volume">vol 9</span> <span class="journal_pages">pp 271-7</span>.</div>
<div class="article_info">R Pilka, G D Norata, H Domanski, C Andersson, S Hansson, P Eriksson and B Casslen. <span class="article_issue_date">2004</span>. <span class="article_title">Matrix metalloproteinase-26 (matrilysin-2) expression is high in endometrial hyperplasia and decreases with loss of histological differentiation in endometrial cancer.</span> <span class="journal_series_title">Gynecol Oncol,</span>, <span class="journal_volume">vol 94</span> <span class="journal_pages">pp 661-70</span>.</div>
<div class="article_info">R Pilka, H Domanski, S Hansson, P Eriksson and B Casslen. <span class="article_issue_date">2004</span>. <span class="article_title">Endometrial TIMP-4 mRNA is high at midcycle and in hyperplasia, but downregulated in malignant tumours. Coordinated expression with MMP-26.</span> <span class="journal_series_title">Mol Hum Reprod,</span>, <span class="journal_volume">vol 10</span> <span class="journal_pages">pp 641-50</span>.</div>
<div class="article_info">R Pilka, I Oborna, V Lichnovsky, P Havelka, H Fingerova, P Eriksson, S Hansson and B Casslén. <span class="article_issue_date"></span>. <span class="article_title">Endometrial expression of the estrogen sensitive genes MMP-26 and TIMP-4 is altered by a substitution protocol without down regulation in IVF patients.</span> <span class="journal_series_title">Hum Reprod</span>, (submitted)</div>
<div class="article_info">R Pilka, V Noskova, H Domanski, C Andersson, S Hansson and B Casslén. <span class="article_issue_date"></span>. <span class="article_title">Endometrial TIMP-4 is derived from the stroma, accumulates in the epithelium, and is released to the uterine fluid.</span> <span class="journal_series_title">Mol Hum Reprod</span>, (submitted)</div>

Ämnesklassifikation (UKÄ)

Reproduktionsmedicin och gynekologi

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@phdthesis{ce83dca889e447949e4226e5cb3f22b6,

title = "Matrix metalloproteinase-26 (MMP-26) and Tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium.",

abstract = "The aim of the thesis was to study matrix metalloproteinase-26 (MMP-26) and tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium. Most importantnly, both these molecules may be ivolved in the implantation process. Trophoblast tissue from human as well as mouse embryos produces pro-MMP-9. Active MMP-9 (gelatinase B) is a proteolytic enzyme with a broad substrate specificity. MMP-26 is an effective activator of pro-MMP-9, and TIMP-4 is a strong inhibitor of MMP-26. We found that endometrial expression of MMP-26 mRNA and TIMP-4 mRNA is elevated during the proliferative phase, is maximal in the early, decreases in the mid-, and is low in the late secretory phases (I, III). The expression pattern of both genes suggests their up-regulation by estrogen, and down-regulation by progesterone. This is supported by our findings of potential estrogen response elements upstream the coding sequences of both genes (II, III). In situ hybridization localised MMP-26 mRNA in epithelial and TIMP-4 mRNA in stromal cells (I, III). As expected, using immunohistochemistry we found MMP-26 protein in the epithelium (II). However, the same technique showed only occasional staining for TIMP-4 in the stroma (V). Instead, TIMP-4 protein was localised in granules of the apical part of epithelial cells. This finding suggests that TIMP-4 is produced in and secreted by the stromal cells, taken up by the epithelial cells, stored in apical granules, and finally secreted to the uterine fluid. Analysis of uterine fluid with Western blot technique demonstrated presence of TIMP-4, but not MMP-26 (V). So far, very few proteins of stromal origin have been demonstrated in the uterine fluid. Furthermore, using different antibodies, we have shown that MMP-26 is stored in epithelial cells in its active form, is not released spontaneously, and when released it is controlled by TIMP-4 in both stroma and uterine fluid. Examination of endometrial tissue obtained from IVF patients in an estrogen and progesterone substitution protocol showed that the estrogen sensitive genes MMP-26 and TIMP-4 are over-stimulated by both estrogen and progesterone. Analysis of MMP-26 and TIMP-4 expression in endometrial hyperplastia showed that both localization and amount of mRNA corresponded to that of the proliferative phase. This is in agreement with estrogen mediated regulation of these genes, as well as hyperplasia being the result of estrogen over-stimulation. The expression of MMP-26 and TIMP-4 decreases with loss of histological differentiation in endometrial cancer. This is also in agreement with the general loss of differentiated functions during progression to poorly differentiated, and more malignant tumors.",

keywords = "cancer, Obstetrics, gynaecology, Cytologi, onkologi, cancerology, secretion, Biology, Biologi, Cytology, oncology, implantation, human, endometrium, mRNA, MMP-26, TIMP-4, reproduktion, andrologi, gynekologi, Obstetrik, sexuality, reproduction, sexualitet, andrology",

author = "Radovan Pilka",

note = "Defence details Date: 2006-04-28 Time: 13:00 Place: Dept. of Obstetrics and Gynecology, Lecture Hall External reviewer(s) Name: Marbaix, Etienne Title: Professor Affiliation: Department of Pathology, University of Louvain Medical School, Brussels, Belgium --- <div class={"}article_info{"}>R Pilka, C Whatling, H Domanski, S Hansson, P Eriksson and B Casslen. <span class={"}article_issue_date{"}>2003</span>. <span class={"}article_title{"}>Epithelial expression of matrix metalloproteinase-26 (MMP-26) is elevated at midcycle in the human endometrium.</span> <span class={"}journal_series_title{"}>Mol Human Reprod,</span>, <span class={"}journal_volume{"}>vol 9</span> <span class={"}journal_pages{"}>pp 271-7</span>.</div> <div class={"}article_info{"}>R Pilka, G D Norata, H Domanski, C Andersson, S Hansson, P Eriksson and B Casslen. <span class={"}article_issue_date{"}>2004</span>. <span class={"}article_title{"}>Matrix metalloproteinase-26 (matrilysin-2) expression is high in endometrial hyperplasia and decreases with loss of histological differentiation in endometrial cancer.</span> <span class={"}journal_series_title{"}>Gynecol Oncol,</span>, <span class={"}journal_volume{"}>vol 94</span> <span class={"}journal_pages{"}>pp 661-70</span>.</div> <div class={"}article_info{"}>R Pilka, H Domanski, S Hansson, P Eriksson and B Casslen. <span class={"}article_issue_date{"}>2004</span>. <span class={"}article_title{"}>Endometrial TIMP-4 mRNA is high at midcycle and in hyperplasia, but downregulated in malignant tumours. Coordinated expression with MMP-26.</span> <span class={"}journal_series_title{"}>Mol Hum Reprod,</span>, <span class={"}journal_volume{"}>vol 10</span> <span class={"}journal_pages{"}>pp 641-50</span>.</div> <div class={"}article_info{"}>R Pilka, I Oborna, V Lichnovsky, P Havelka, H Fingerova, P Eriksson, S Hansson and B Cassl{\'e}n. <span class={"}article_issue_date{"}></span>. <span class={"}article_title{"}>Endometrial expression of the estrogen sensitive genes MMP-26 and TIMP-4 is altered by a substitution protocol without down regulation in IVF patients.</span> <span class={"}journal_series_title{"}>Hum Reprod</span>, (submitted)</div> <div class={"}article_info{"}>R Pilka, V Noskova, H Domanski, C Andersson, S Hansson and B Cassl{\'e}n. <span class={"}article_issue_date{"}></span>. <span class={"}article_title{"}>Endometrial TIMP-4 is derived from the stroma, accumulates in the epithelium, and is released to the uterine fluid.</span> <span class={"}journal_series_title{"}>Mol Hum Reprod</span>, (submitted)</div>",

year = "2006",

language = "English",

isbn = "91-85481-73-4",

publisher = "Publishing Centre, Palacky University, Olomouc, Czech Republic",

type = "Doctoral Thesis (compilation)",

school = "Obstetrics and Gynaecology (Lund)",

}

TY - THES

T1 - Matrix metalloproteinase-26 (MMP-26) and Tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium.

AU - Pilka, Radovan

N1 - Defence details Date: 2006-04-28 Time: 13:00 Place: Dept. of Obstetrics and Gynecology, Lecture Hall External reviewer(s) Name: Marbaix, Etienne Title: Professor Affiliation: Department of Pathology, University of Louvain Medical School, Brussels, Belgium --- <div class="article_info">R Pilka, C Whatling, H Domanski, S Hansson, P Eriksson and B Casslen. <span class="article_issue_date">2003</span>. <span class="article_title">Epithelial expression of matrix metalloproteinase-26 (MMP-26) is elevated at midcycle in the human endometrium.</span> <span class="journal_series_title">Mol Human Reprod,</span>, <span class="journal_volume">vol 9</span> <span class="journal_pages">pp 271-7</span>.</div> <div class="article_info">R Pilka, G D Norata, H Domanski, C Andersson, S Hansson, P Eriksson and B Casslen. <span class="article_issue_date">2004</span>. <span class="article_title">Matrix metalloproteinase-26 (matrilysin-2) expression is high in endometrial hyperplasia and decreases with loss of histological differentiation in endometrial cancer.</span> <span class="journal_series_title">Gynecol Oncol,</span>, <span class="journal_volume">vol 94</span> <span class="journal_pages">pp 661-70</span>.</div> <div class="article_info">R Pilka, H Domanski, S Hansson, P Eriksson and B Casslen. <span class="article_issue_date">2004</span>. <span class="article_title">Endometrial TIMP-4 mRNA is high at midcycle and in hyperplasia, but downregulated in malignant tumours. Coordinated expression with MMP-26.</span> <span class="journal_series_title">Mol Hum Reprod,</span>, <span class="journal_volume">vol 10</span> <span class="journal_pages">pp 641-50</span>.</div> <div class="article_info">R Pilka, I Oborna, V Lichnovsky, P Havelka, H Fingerova, P Eriksson, S Hansson and B Casslén. <span class="article_issue_date"></span>. <span class="article_title">Endometrial expression of the estrogen sensitive genes MMP-26 and TIMP-4 is altered by a substitution protocol without down regulation in IVF patients.</span> <span class="journal_series_title">Hum Reprod</span>, (submitted)</div> <div class="article_info">R Pilka, V Noskova, H Domanski, C Andersson, S Hansson and B Casslén. <span class="article_issue_date"></span>. <span class="article_title">Endometrial TIMP-4 is derived from the stroma, accumulates in the epithelium, and is released to the uterine fluid.</span> <span class="journal_series_title">Mol Hum Reprod</span>, (submitted)</div>

PY - 2006

Y1 - 2006

N2 - The aim of the thesis was to study matrix metalloproteinase-26 (MMP-26) and tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium. Most importantnly, both these molecules may be ivolved in the implantation process. Trophoblast tissue from human as well as mouse embryos produces pro-MMP-9. Active MMP-9 (gelatinase B) is a proteolytic enzyme with a broad substrate specificity. MMP-26 is an effective activator of pro-MMP-9, and TIMP-4 is a strong inhibitor of MMP-26. We found that endometrial expression of MMP-26 mRNA and TIMP-4 mRNA is elevated during the proliferative phase, is maximal in the early, decreases in the mid-, and is low in the late secretory phases (I, III). The expression pattern of both genes suggests their up-regulation by estrogen, and down-regulation by progesterone. This is supported by our findings of potential estrogen response elements upstream the coding sequences of both genes (II, III). In situ hybridization localised MMP-26 mRNA in epithelial and TIMP-4 mRNA in stromal cells (I, III). As expected, using immunohistochemistry we found MMP-26 protein in the epithelium (II). However, the same technique showed only occasional staining for TIMP-4 in the stroma (V). Instead, TIMP-4 protein was localised in granules of the apical part of epithelial cells. This finding suggests that TIMP-4 is produced in and secreted by the stromal cells, taken up by the epithelial cells, stored in apical granules, and finally secreted to the uterine fluid. Analysis of uterine fluid with Western blot technique demonstrated presence of TIMP-4, but not MMP-26 (V). So far, very few proteins of stromal origin have been demonstrated in the uterine fluid. Furthermore, using different antibodies, we have shown that MMP-26 is stored in epithelial cells in its active form, is not released spontaneously, and when released it is controlled by TIMP-4 in both stroma and uterine fluid. Examination of endometrial tissue obtained from IVF patients in an estrogen and progesterone substitution protocol showed that the estrogen sensitive genes MMP-26 and TIMP-4 are over-stimulated by both estrogen and progesterone. Analysis of MMP-26 and TIMP-4 expression in endometrial hyperplastia showed that both localization and amount of mRNA corresponded to that of the proliferative phase. This is in agreement with estrogen mediated regulation of these genes, as well as hyperplasia being the result of estrogen over-stimulation. The expression of MMP-26 and TIMP-4 decreases with loss of histological differentiation in endometrial cancer. This is also in agreement with the general loss of differentiated functions during progression to poorly differentiated, and more malignant tumors.

AB - The aim of the thesis was to study matrix metalloproteinase-26 (MMP-26) and tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium. Most importantnly, both these molecules may be ivolved in the implantation process. Trophoblast tissue from human as well as mouse embryos produces pro-MMP-9. Active MMP-9 (gelatinase B) is a proteolytic enzyme with a broad substrate specificity. MMP-26 is an effective activator of pro-MMP-9, and TIMP-4 is a strong inhibitor of MMP-26. We found that endometrial expression of MMP-26 mRNA and TIMP-4 mRNA is elevated during the proliferative phase, is maximal in the early, decreases in the mid-, and is low in the late secretory phases (I, III). The expression pattern of both genes suggests their up-regulation by estrogen, and down-regulation by progesterone. This is supported by our findings of potential estrogen response elements upstream the coding sequences of both genes (II, III). In situ hybridization localised MMP-26 mRNA in epithelial and TIMP-4 mRNA in stromal cells (I, III). As expected, using immunohistochemistry we found MMP-26 protein in the epithelium (II). However, the same technique showed only occasional staining for TIMP-4 in the stroma (V). Instead, TIMP-4 protein was localised in granules of the apical part of epithelial cells. This finding suggests that TIMP-4 is produced in and secreted by the stromal cells, taken up by the epithelial cells, stored in apical granules, and finally secreted to the uterine fluid. Analysis of uterine fluid with Western blot technique demonstrated presence of TIMP-4, but not MMP-26 (V). So far, very few proteins of stromal origin have been demonstrated in the uterine fluid. Furthermore, using different antibodies, we have shown that MMP-26 is stored in epithelial cells in its active form, is not released spontaneously, and when released it is controlled by TIMP-4 in both stroma and uterine fluid. Examination of endometrial tissue obtained from IVF patients in an estrogen and progesterone substitution protocol showed that the estrogen sensitive genes MMP-26 and TIMP-4 are over-stimulated by both estrogen and progesterone. Analysis of MMP-26 and TIMP-4 expression in endometrial hyperplastia showed that both localization and amount of mRNA corresponded to that of the proliferative phase. This is in agreement with estrogen mediated regulation of these genes, as well as hyperplasia being the result of estrogen over-stimulation. The expression of MMP-26 and TIMP-4 decreases with loss of histological differentiation in endometrial cancer. This is also in agreement with the general loss of differentiated functions during progression to poorly differentiated, and more malignant tumors.

KW - cancer

KW - Obstetrics

KW - gynaecology

KW - Cytologi

KW - onkologi

KW - cancerology

KW - secretion

KW - Biology

KW - Biologi

KW - Cytology

KW - oncology

KW - implantation

KW - human

KW - endometrium

KW - mRNA

KW - MMP-26

KW - TIMP-4

KW - reproduktion

KW - andrologi

KW - gynekologi

KW - Obstetrik

KW - sexuality

KW - reproduction

KW - sexualitet

KW - andrology

M3 - Doctoral Thesis (compilation)

SN - 91-85481-73-4

PB - Publishing Centre, Palacky University, Olomouc, Czech Republic

ER -

Matrix metalloproteinase-26 (MMP-26) and Tissue inhibitor of metalloproteinases-4 (TIMP-4) in normal, hyperplastic, and malignant endometrium.

Sammanfattning

Bibliografisk information

Ämnesklassifikation (UKÄ)

FN:s Globala mål

Fingeravtryck

Citera det här