Evaluation of Activated Leukocyte Cell Adhesion Molecule as a Biomarker for Breast Cancer in ...

153 Pages·2013·4.42 MB·English

Evaluation of Activated Leukocyte Cell
Adhesion Molecule as a Biomarker for
Breast Cancer in Egyptian Patients

Presented by

Mostafa Saif El-Nasr Mahmoud El-Shepiny
M. Sc. in Biochemistry
Assistant Lecturer in National Center for Radiation
Research and Technology
Atomic Energy Authority

A Thesis Submitted
to
Faculty of Science

In Partial Fulfillment of the
Requirements for the Degree of
Ph. D. of Science
(Biochemistry)

Under Supervision of

Prof. Dr. Amr Saad Mohammed Prof. Dr. Azza Abd-Alla Mohammed
Professor of Biochemistry Professor of Clinical Pathology
Faculty of Science National Center for Radiation Research
Cairo University and Technology
Atomic Energy Authority
Prof. Dr. Amal Mohammed Nour El-Din Dr. Ahmed Mostafa Ahmed
Professor of Pediatric Medicine Lecturer of Surgical Oncology
Nuclear Research Center National Cancer Institute
Atomic Energy Authority Cairo University

Chemistry Department
Faculty of Science
Cairo University

(2013)
Approval Sheet for Submission
Title of (Ph. D.) thesis: Evaluation of Activated Leukocyte Cell Adhesion
Molecule as a Biomarker for Breast Cancer in Egyptian Patients
Name of the candidate: Mostafa Saif El-Nasr Mahmoud El-Shepiny
This thesis has been approved for submission by the supervisors:
1. Prof. Dr. Amr Saad Mohammed
Professor of Biochemistry
Faculty of Science
Cairo University
2. Prof. Dr. Azza Abd-Alla Mohammed
Professor of Clinical Pathology
National Center for Radiation Research and Technology
Atomic Energy Authority
3. Prof. Dr. Amal Mohammed Nour El-Din
Professor of Pediatric Medicine
Nuclear Research Center
Atomic Energy Authority
4. Dr. Ahmed Mostafa Ahmed
Lecturer of Surgical Oncology
National Cancer Institute
Cairo University

Prof. Dr. Hamed Abd El-Latif Abd El-Rahman

Chairman of Chemistry Department
Faculty of Science - Cairo University

Abstract

Abstract
Name: Mostafa Saif El-Nasr Mahmoud El-Shepiny
Title of thesis: Evaluation of Activated Leukocyte Cell Adhesion Molecule as a
Biomarker for Breast Cancer in Egyptian Patients
Degree: (Ph. D.) thesis, Faculty of Science, Cairo University, 2013
In this study, serum activated leukocyte cell adhesion molecule (ALCAM)
levels were evaluated in 41 primary breast cancer patients and 20 healthy
females, and its diagnostic value was quantified, and compared with those of
carbohydrate antigen 15-3 (CA15-3) and carcinoembryonic antigen (CEA).
Also, its prognostic value was examined. Serum ALCAM levels were also
evaluated before and after surgical treatment. Serum levels of ALCAM and CA
15-3 were significantly higher in breast cancer patients than healthy controls
(P=0.002, P=0.043 respectively), but the difference in serum CEA levels did not
reach statistical significance. Serum ALCAM levels had significant area under
the curve (AUC) (P=0.002), but serum levels of CA 15-3 and CEA had non-
significant AUCs, and various combinations between them did not result in any
improvement. A significant association was found between serum levels of
ALCAM and CEA with age and menopausal status in breast cancer patients.
Non-significant difference was shown in serum levels of ALCAM, CA 15-3 and
CEA before and after surgical treatment. In conclusion, this study suggests that
serum ALCAM may represent a novel diagnostic biomarker for breast cancer.
Key words: ALCAM, Breast cancer, CA15-3, CEA, and biomarker.
Supervisors:
Prof. Dr. Amr Saad Mohammed Prof. Dr. Azza Abd-Alla Mohammed
Prof. Dr. Amal Mohammed Nour El-Din Dr. Ahmed Mostafa Ahmed
Prof. Dr. Hamed Abd El-Latif Abd El-Rahman
Chairman of Chemistry Department
Faculty of Science - Cairo University
Acknowledgement

Acknowledgement
Thanks to Allah first and foremost. I feel always indebted to Allah the
kindest and the most beneficent and merciful.
I would like to express my great thanks and deepest appreciation to Prof.
Dr. Amr Saad Mohammed, professor of biochemistry, Faculty of Science,
Cairo University, for his kind supervision, valuable guidance, support,
invaluable advices and great help.
My grateful acknowledgement, great thanks, sincere appreciation and
deepest respect to Prof. Dr. Azza Abd-Alla Mohammed, professor of clinical
pathology, National Center for Radiation Research and Technology, Atomic
Energy Authority, for her tremendous effort, precious guidance, helpful
instructions and powerful support under her continuous kind supervision to
finish this study.
I would like to express my great thanks to Prof. Dr. Amal Mohammed
Nour El-Din, professor of pediatric medicine, Nuclear Research Center, Atomic
Energy Authority,, for her valuable advices, support and continuous
encouragement.
My great thanks and gratitude to Dr. Ahmed Mostafa Ahmed, lecturer
of surgical oncology, National Cancer Institute, Cairo University, for his kind
supervision, valuable guidance, advices and help in obtaining patients samples
and their clinical data.
I would like to thank all subjects who participated in this study.
Finally, my deepest thanks to all my family members for their
continuous help and encouragement especially my brother Ahmed.
To my beloved mother
To my brothers
and my sister
Contents

Contents

Page
List of abbreviations…………………………………………………… I
List of tables…………………………………………………………….. V
List of figures…………………………………………………………… VI
Introduction…………………………………………………………….. 1
Aim of the work………………………………………………………… 4
Review of literature…………………………………………………….. 5
Breast cancer………………………………………………………. 5
Breast cancer risk factors…………………………………… 5
Staging of breast cancer…………………………………….. 21
World Health Organization classification of breast tumor…. 24
Histopathological grades of breast cancer…………………... 26
Serum tumor markers in breast cancer……………………… 26
Cell adhesion molecules.………………………………………….. 36
Integrins.…………………………………………………….. 36
Cadherins……………………………………………………. 37
Selectins…………………………………………………….. 38
Immunoglobulin-like cell adhesion molecules……………… 39
Activated leukocyte cell adhesion molecule.……………………… 42
Transcriptional regulation and function.……………………. 42
Activated leukocyte cell adhesion molecule and various
types of cancer.……………………………………………… 44
Subjects and methods………………………………………………….. 52
Subjects……………………………………………………………. 52
Specimen collection.………………………………………………. 54
Determination of activated leukocyte cell adhesion molecule.…… 55
Determination of carbohydrate antigen 15-3……………………… 59
Contents

Determination of carcinoembryonic antigen.……………………... 61
Determination of aspartate aminotransferase.…………………….. 63
Determination of alanine aminotransferase.………………………. 64
Determination of urea.…………………………………………….. 64
Determination of creatinine.………………………………………. 65
Statistical analysis.………………………………………………… 65
Results…………………………………………………………………... 67
Discussion……………………………………………………………….. 85
Summary………………………………………………………………... 90
References………………………………………………………………. 94
Arabic summary
List of Abbreviations

List of Abbreviations
A Adenine
ADAM(s) A disintegrin and metalloproteinase(s)
ADH Alcohol dehydrogenase
AHH Aryl hydrocarbon hydroxylase
AICR American Institute for Cancer Research
ALCAM Activated leukocyte cell adhesion molecule
ALT Alanine aminotransferase
AP-1 Activator protein-1
ASCO American Society of Clinical Oncology
AST Aspartate aminotransferase
AT Ataxia telangiectasia
ATM Ataxia telangiectasia mutated
AUC(s) Area under the curve(s)
bp Base pair
BRCA Breast cancer
C Cytosine
CA Carbohydrate antigen
CAM(s) Cell adhesion molecule(s)
CARE Contraceptive and Reproductive Experiences
CD Cluster of differentiation
CEA Carcinoembryonic antigen
CI Confidence interval
COMA Committee on Medical Aspects of Food and Nutrition Policy
CPM Counts per minute
CYP Cytochrome P450
DNA Deoxyribonucleic acid
I
List of Abbreviations

EC Extracellular cadherin
ECM Extracellular matrix
EGF Epidermal growth factor
EGTM European Group on Tumor Markers
ELISA Enzyme-linked immunosorbent assay
ER Estrogen receptor
ESMO European Society of Medical Oncology
FA Fanconi anemia
FAK Focal adhesion kinase
Fc Fragment crystallizable
FN Fibronectin
Fra-2 Fos-related antigen-2
G Guanine
GFP Green fluorescent protein
GH Growth hormone
GSH Glutathione
GST Glutathione S-transferase
HB2 High density lipoprotein-binding protein 2
HER-2 Human epidermal growth factor receptor-2
HEV(s) High endothelial venule(s)
HRP Horseradish peroxidase
HRT Hormone replacement therapy
HSP70 Heat shock protein 70
ICAM(s) Intercellular adhesion molecule(s)
Ig Immunoglobulin
Ig-CAM(s) Immunoglobulin-like cell adhesion molecule(s)
IGF Insulin-like growth factor
IGFBP-3 Insulin-like growth factor binding protein-3
II