Polymorphism and expression of IL-10 in serum and ascites from patients with advanced ovarian cancer [Elektronische Ressource] / von Elena Ioana Braicu

Ioana

Sujets

• Gesundheit

Informations

• Publié par: charite_-_universitatsmedizin_berlin
• Publié le: 01 janvier 2011
• Nombre de lectures: 55
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Aus der Klinik für Gynäkologie/Campus Virchow Klinikum
der Medizinischen Fakultät Charité – Universitätsmedizin Berlin



DISSERTATION


Polymorphism and expression of IL-10 in serum and ascites
from patients with advanced ovarian cancer



zur Erlangung des akademischen Grades
Doctor medicinae (Dr. med.)







vorgelegt der Medizinischen Fakultät
Charité – Universitätsmedizin Berlin





von


Elena-Ioana Braicu

aus Cluj-Napoca


































Gutachter/in: 1. Prof. Dr. med. J. Sehouli
2. Prof. Dr. med. C. Denkert
3. Prof. Dr. med. W. Friedmann


Datum der Promotion: 04.02.2011



2 Contents
1 Introduction ............................................................................................................... 5
1.1 Ovarian cancer .. 5
1.1.1 Epidemiology ................................................................................................. 7
1.1.2 Etiology and risk factors ................ 8
1.1.3 Histological classification of epithelial tumors ............................................. 13
1.1.4 Grading ....... 16
1.1.5 The natural history of ovarian cancer .......................... 16
1.1.6 FIGO and TNM classifications .................................... 16
1.1.7 Prognostic factors ....................................................... 17
1.1.8 Screening .................................... 22
1.1.9 Diagnostics .. 24
1.1.10 Therapy ....................................................................... 24
1.2 Interleukin-10 involvement in tumor development ........... 27
1.2.1 Cancer immunology .................................................... 27
1.2.2 The cytokines network ................ 30
1.2.3 Structure and function ................................................. 31
1.2.4 Interleukin-10 and cancer ............ 33
1.2.5 Interleukin-10 polymorphism ....................................... 35
1.2.6 Interleukin-10 and ovarian cancer ............................................................... 38
1.3 Study objectives ................................ 39
2 Methods .................................................. 41
2.1 The collective of patients ................................................................................. 41
2.2 Collection of blood and ascites samples ......................... 42
2.3 Enzyme-Linked Immunosorbent Assay (ELISA) .............. 42
2.4 DNA extraction ................................................................................................. 47
2.5 Determining the DNA concentration using UV-spectrophotometry ................. 48
2.6 Polymerase Chain Reaction (PCR) . 49
2.7 Pyrosequencing ............................... 52
2.8 Statistics .......................................................................................................... 56
3 Results .................... 58
3.1 IL-10 expression in serum and ascites ............................................................ 58
3.1.1 Patients characteristics ............... 58
3.1.2 Levels of IL-10 in serum and ascites in the ovarian cancer and control
groups ................................................................................................................. 65
3.1.3 Expression of IL-10 in serum and ascites in patients with ovarian cancer .. 67
3.1.4 Correlation between established clinical prognostic factors and the
expression of IL-10 ................................................................................................. 68
3.1.5 The predictive value of IL-10 expression in serum and ascites for patients
with ovarian cancer . 72
3.1.6 The impact of IL-10 expression in serum and ascites on overall and
tumor-free survival rates in patients with ovarian cancer ........................................ 73
3.2 IL-10 polymorphisms ....................................................... 75
3.2.1 Patients’ characteristics .............................................. 75
3.2.2 The IL-10 promoter polymorphism in ovarian cancer group 85
3.2.3 IL-10 promoter polymorphism in ovarian cancer patients and in the
control group .......................................................................... 87
3.2.4 Correlation between IL-10 promoter polymorphism and the established
prognostic factors ................................... 88
3 3.2.5 Correlation between IL-10 promoter polymorphism and the residual
tumor mass and overall survival ............................................................................ 91
3.3 Correlation between the expression and the polymorphism of IL-10 in
patients with ovarian cancer ....................... 96
3.3.1 Patients’ characteristics .............. 96
3.3.2 Impact of IL-10 polymorphisms on the expression in serum and ascites ... 98
3.4 The impact of tumor debulking on the expression of IL-10 ............................ 104
3.4.1 Patients’ characteristics ............................................................................ 104
3.4.2 ELISA ........................................ 105
4 Discussion ............ 111
4.1 The overexpression of IL-10 in ovarian cancer patients ................................ 111
4.2 The IL-10 promoter gene polymorphism ....................................................... 115
4.3 Does the allele frequency have an impact on the expression of IL-10? ........ 117
5 Conclusions .......................................................................... 119
6 Abstract ................. 120
7 Literature ............................................... 124
8 Summary of the tables and figures ....... 138
8.1 Tables ............................................................................ 138
8.2 Figures ........... 139
9 List of abbreviations .............................................................................................. 141
10 Acknowledgement ................................ 142
11 Curriculum Vitae ... 143
11.1 Personal Details ................. Error! Bookmark not defined.
11.2 Contact details ................................................... Error! Bookmark not defined.
11.3 Professional Experience .... Error! Bookmark not defined.
11.4 Education ........................... Error! Bookmark not defined.
11.5 Prizes, Awards and Scholarships ...................... Error! Bookmark not defined.
11.6 Languages ......................................................... Error! Bookmark not defined.
12 List of publications ................................................................ 144
12.1 Book Chapters ............................... 144
12.2 Journal Articles .............................. 144
12.3 Abstracts, posters and conference proceedings ............................................ 145
13 Erklärung der Eigenständigkeit ............................................. 147

4 IL-10 Expression and Polymorphism in Ovarian cancer Introduction
1 Introduction

1.1 Ovarian cancer

thOvarian cancer is the 5 most frequent malignant disease of the European
woman. Due to poor prognosis, it is the leading cause of death related to gynecologic
tumors (1). In Germany, about 8,000 women are diagnosed every year as having a
malignant ovarian tumor. Because no effective screening method is available until now
and due to the poor clinical behavior most of the patients are diagnosed in FIGO stage
III or IV (1, 2).
In Europe the highest incidence of ovarian cancer is in the Scandinavian
countries, the lowest one being in the south European countries.
As screening methods bimanual pelvic examination, ultrasonography and
measurement of the CA-125 serum levels were investigated in large trials.
Unfortunately the methods are not specific and sensitive enough to fulfill the
requirements for an efficient screening test (3, 4, 5). The new study presented at ASCO
in 2009, which included 1442 patients from 10 countries, was designed in order to
determine the benefits of early treatment based only on the elevation of CA-125
compared to delayed treatment until clinically indicated. Although the treatment was
started 5 months earlier in the immediate arm, the study showed no significant impact
on the overall survival between the two arms (hazard ratio 1.01, 95% CI 0.82-1.25, p =
0.91) (6).
Oskay-Oezcelik et al. presented at ASCO the results from a multi-institutional
survey about cancer care in ovarian cancer patients. The study included 1060 patients.
According the patients’ opinion, the main objective for the follow-up is the early
detection of relapse and a prolongation of overall survival (95.8%). More than 90% get
CA-125 measurements, being the procedure with highest anxiety but also the most
important procedure for the patient (7).
CA-125 antigen is a useful serum mark