LIST OF CONTENTS

Title Page No
Declaration i
Certificate ii
Acknowledgement iii
List of contents v
List of tables vii
List of figures viii
List of abbreviations ix
Abstract xi
Chapter 1. INTRODUCTION
1.1. Background 1
1.2. Rationale 5
1.3. Research Question 6
1.4. Objectives 7
Chapter 2. REVIEW OF LITERATURES 8-23
Chapter 3. MATERIALS AND METHODS 24-31
3.1. Type of study 24
3.2. Study Period 24
3.3. Study Population 24
3.4. Place of study 24
3.5. Sampling technique 24
3.6. Selection criteria 25
3.6.1. Inclusion criteria 26
3.6.2. Exclusion criteria 26
3.7. Sample size determination 26
3.8. Sample collection and processing 28
3.8.1. History of the patient 28
3.8.2. Histopathology 28
3.8.3. Immunohistochemistry 28
3.8.4. Assessment of WT1 29
3.8.5. Statistical analysis. 30
3.9 Ethical consideration 31

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Chapter 4. RESULTS AND OBSERVATION 32-39
Chapter 5. DISCUSSION 40-44
Chapter 6. SUMMARY, CONCLUSION, LIMITATION AND 45-48
RECOMMENDATIONS
6.1. Summary 45
6.2. Conclusion 46
6.3. Limitation 47
6.4. Recommendation 48
REFERENCES 49-56
APPENDICES xiii-xxxviii
I. Informed Written Consent (English) xiii
I. Consent form xiv
II. Data Collection Sheet xvi
III. Processing of Tissue for paraffin impregnation xviii
IV. Preparation of stains and chemicals for H & E stain xxiii
V. Immunohistochemistry staining procedure xxvi
VI. Master Table xxviii
VII. IRB certificate xxx
VIII. Ethical clearance certificate xxxi
VIII. Gross & microscopic picture xxxii

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 LIST OF TABLES
Table No Title Page No

Table I Clinical and molecular differences between main histologic 19

subtypes of ovarian carcinoma.

Table II Distribution of subject according to age. 32

Table III Distribution of WT1 scores. 34

Table IV Distribution of WT1 scores in different histologic subtypes. 35

Table V WT1 expression in different histologic subtypes of ovarian 36

carcinoma.

Table VI Relation of WT1 scores with age of the subject. 37

Table VII WT1 literature data vs. personal results. 39

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 LIST OF FIGURES

Fig No Title Page no

Figure 1 Pie chart Showing distribution of histologic subtypes. 33

Figure 2 Bar diagram showing frequency of expression of WT1 in 38

different histologic subtypes.

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 LIST OF ABBREVIATIONS

AIHW Australia Institute of Health and Welfare

AJCC American Joint Committee on Cancer

ARID1A AT-Rich Interaction Domain 1A

BRCA BReast CAncer gene

BSMMU Bangabandhu Sheikh Mujib Medical University

BOCS Breast Ovarian Cancer Syndrome

CCC Clear Cell Carcinoma

CA-125 Cancer Antigen 125

EC Endometrioid Carcinoma

H&E Haematoxylin and Eosin

HER-2 Human Epidermal Growth Factor Receptor-2

HPF High Power Field

HNPCCS Hereditary Non Polyposis Colorectal Cancer Syndrome

HGSC High Grade Serous Carcinoma

IHC Immunohistochemistry

IRB Institutional Review Board

KRAS Kirsten RAt Sarcoma virus

LGSC Low Grade Serous Carcinoma

LOH Loss of Heterozygosity

MOC Mucinous Ovarian Carcinoma

NBCC National Breast Cancer Center

PAX 8 Paired Box Gene 8

PI3K Phosphoinositide 3-Kinase

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PTEN Phosphatase and TENsin homolog

RMC Rajshahi Medical College

RMCH Rajshahi Medical College Hospital

SD Standard Deviation

SEER Surveillance, Epidemiology and End Result Program

SEM Standard Error of mean

SOC Serous Ovarian Carcinoma

SPSS Statistical Package for Social Sciences

SSOCS Site Specific Ovarian Cancer Syndrome

TP53 Tumor Protein 53

TSG Tumor Suppressor Gene

USA United States of America

WHO World Health Organization.

Wnt Wingless related integration site

WT1 Wilms Tumor Gene 1

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 ABSTRACT

Ovarian carcinoma is the seventh most common cancer among women worldwide and

eighth leading cause of death. Epithelial ovarian carcinoma is the most common type and

consists of 60%-70% among all types of ovarian cancer. The aim of the present study

was to evaluate variations in Wilms Tumour Gene (WT1) expression among different

histologic subtypes of ovarian carcinomas. WT1 was originally identified as a tumor

suppressor gene located on chromosome 11p13. It has distinct roles in organogenesis,

cells growth and differentiation. Recently, their expression was also confirmed in a

variety of malignancies, being included in the antibodies panel recommended for the

female genital tract pathology. This cross sectional type of descriptive study was

conducted in the Department of Pathology, Rajshahi Medical College (RMC) over a

period of two years from July 2019 to June 2021. A total number of 31 clinically

suspected cases of ovarian carcinoma admitted in the Rajshahi Medical College Hospital

(RMCH) and later on histopathologically confirmed as ovarian carcinoma were included

in the study. History of the patients were obtained from hospital records. One

representative formalin-fixed and paraffin-embedded tissue block was selected. One

slide from each case included in the study was immunostained using the WT1 clone 6F-

H2. The immunoreactivity was graded according to the percentage of stained tumor

cells. Only nuclear staining was considered a positive reaction. A tumor was regarded as

negative if less than 1% of the tumor cells was stained. All serous carcinoma (16/16)

showed WT1 expression, whereas all mucinous (4/4) and all clear cell carcinoma (3/3)

were negative. Two (2) undifferentiated carcinoma included in the study were also

negative. Four (4) (66.6%) of six (6) endometrioid carcinomas were negative whereas

two (2) (33.33%) endometrioid carcinomas showed weak & medium positive expression.

The present study demonstrates differences in immunohistochemical expression of WT1

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among different histologic subtypes of ovarian carcinoma. Additional studies on

endometrioid carcinoma are needed. Another study may be done to find out the

prognostic role of WT1 in ovarian carcinoma. Our study provide supplementary

evidences to support the association of WT1 immunostaining in the investigation of the

ovarian carcinoma. In our country, it is recommended for clinician to use WT1 as an

effective marker in the differential diagnosis of epithelial ovarian carcinoma. Patient may

also be benefited as it guide a physician for taking his or her further approach regarding

therapeutic management. So, it could be concluded that, in addition to histopathological

diagnosis, WT1 could be used as an essential marker to differentiate histologic subtypes

of ovarian carcinoma.

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