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Open Access Research article

Fibroid explants reveal a higher sensitivity against MDM2-inhibitor nutlin-3 than matching myometrium

Dominique N Markowski1, Burkhard M Helmke2, Arlo Radtke1, Jennifer Froeb1, Gazanfer Belge1, Sabine Bartnitzke1, Werner Wosniok3, Iris Czybulka-Jachertz4, Ulrich Deichert5 and Jörn Bullerdiek16*

Author Affiliations

1 Center of Human Genetics, University of Bremen, Leobener Strasse ZHG, D-28359 Bremen, Germany

2 Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 220/221, D-69120 Heidelberg, Germany, current address of B.M.H.: Institute of Pathology, Elbe Kliniken, Klinikum Stade, Bremervörder Str. 111, D- 21682 Stade, Germany

3 Institute of Statistics, University of Bremen, Am Fallturm 1, 28359 Bremen, Germany

4 Clinic of Gynecology and Obstetrics, Elbe Kliniken, Klinikum Stade, Bremervörder Str. 111, 21682 Stade, Germany

5 Clinic of Gynecology and Obstetrics, Krankenhaus Cuxhaven, Altenwalder Chaussee 10, D-27474 Cuxhaven, Germany

6 Small Animal Clinic, University of Veterinary Medicine, Bünteweg 9, D-30559 Hannover, Germany

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BMC Women's Health 2012, 12:2  doi:10.1186/1472-6874-12-2

Published: 10 January 2012

Abstract

Background

Spontaneous cessation of growth is a frequent finding in uterine fibroids. Increasing evidence suggests an important role of cellular senescence in this growth control. Deciphering the underlying mechanisms of growth control that can be expected not only to shed light on the biology of the tumors but also to identify novel therapeutic targets.

Methods

We have analyzed uterine leiomyomas and matching normal tissue for the expression of p14Arf and used explants to see if reducing the MDM2 activity using the small-molecule inhibitor nutlin-3 can induce p53 and activate genes involved in senescence and/or apoptosis. For these studies quantitative real-time RT-PCR, Western blots, and immunohistochemistry were used. Statistical analyses were performed using the student's t test.

Results

An in depth analysis of 52 fibroids along with matching myometrium from 31 patients revealed in almost all cases a higher expression of p14Arf in the tumors than in the matching normal tissue. In tissue explants, treatment with the MDM2 inhibitor nutlin-3 induced apoptosis as well as senescence as revealed by a dose-dependent increase of the expression of BAX as well as of p21, respectively. Simultaneously, the expression of the proliferation marker Ki-67 drastically decreased. Western-blot analysis identified an increase of the p53 level as the most likely reason for the increased activity of its downstream markers BAX and p21. Because as a rule fibroids express much higher levels of p14Arf, a major negative regulator of MDM2, than matching myometrium it was then analyzed if fibroids are more sensitive against nutlin-3 treatment than matching myometrium. We were able to show that in most fibroids analyzed a higher sensibility than that of matching myometrium was noted with a corresponding increase of the p53 immunopositivity of the fibroid samples compared to those from myometrium.

Conclusions

The results show that uterine fibroids represent a cell population of advanced cellular age compared to matching myometrium. Moreover, the data point to members of the p53-network as to potential novel therapeutic targets for the treatment of uterine fibroids.

Keywords:
fibroids; p14Arf; senescence; apoptosis; MDM2 antagonists