characteristics associated with normal stem cells, specifically the ability to give ... I would like to express my sincere gratitude to my supervisor ... words to express my gratitude and never be able to ... To my dearest brothers who without their.
Immunohistochemical Assessment of Cancer Stem Cells in Mucoepidermoid Carcinoma Omnia Ibrahim Afifi*, Samia Mostafa El-Azab** and Rehab Fawzi Kasem*** Abstract Background: Cancer stem cells (CSCs) are cancer cells that possess characteristics associated with normal stem cells, specifically the ability to give rise to all cell types found in a particular cancer sample. Such cells are proposed to persist in tumors as a distinct population and cause relapse and metastasis by giving rise to new tumors. Therefore, development of specific therapies targeted at CSCs holds hope for improvement of survival and quality of life of cancer patients. CD133 was reported as a putative marker for identification of CSC in different tumors. However, no studies investigated the presence of CSCs in mucoepidermoid carcinoma (MEC). Materials and Methods: The presence of CSCs in paraffin embedded tissue sections of twelve MEC cases (6 low grade and 6 high grade) was investigated by the expression of CD133 immunohistochemically. Identification and counting of CD133 positive colonies of cells was done using a light microscope. Welixon test was used to detect the statistical significance and to compare CD133 positivity between low grade and high grade MEC. Results: Low grade MEC showed moderate and many CD133 + colonies of cells. Whereas high grade cases showed negative, few and moderate CD133+ cells. Conclusion: The current detection of CD133+ colonies in MEC suggests that MEC follows CSCs model, the CSCs count decrease with increasing the grade of the carcinoma. Therefore, these cells may play a role in tumor initiation. Additionally CD133 is a potential cancer stem cell marker in MEC. Key Words: Cancer Stem Cells, Mucoepidermoid Carcinoma, Salivary gland tumors, CD133
ﺗﻘﯿﯿﻢ اﻟﻤﻨﺎﻋﺔ اﻟﻨﺴﯿﺠﯿﺔ اﻟﻜﯿﻤﯿﺎﺋﯿﺔ ﻟﻠﺨﻼﯾﺎ اﻟﺠﺬﻋﯿﺔ اﻟﺴﺮطﺎﻧﯿﺔ ﻓﻰ ﺳﺮطﺎن اﻟﺨﻼﯾﺎ اﻟﻤﺨﺎطﯿﺔ اﻟﺒﺸﺮاﻧﯿﺔ
رﺳﺎﻟﺔ ﻣﻘﺪﻣﺔ إﻟﻰ ﻛﻠﯿﺔ طﺐ اﻟﻔﻢ واﻷﺳﻨﺎن ﺟﺎﻣﻌﺔ اﻟﻘﺎھﺮة ﻛﺠﺰء ﺗﻜﻤﯿﻠﻲ ﻟﻤﺘﻄﻠﺒﺎت اﻟﺤﺼﻮل ﻋﻠﻰ درﺟﺔ اﻟﻤﺎﺟﯿﺴﺘﯿﺮ ﻓﻲ اﻟﻌﻠﻮم اﻷﺳﺎﺳﯿﺔ ﻟﻄﺐ اﻷﺳﻨﺎن )ﺑﺎﺛﻮﻟﻮﺟﯿﺎ اﻟﻔﻢ(
ﻣﻘﺪﻣﺔ ﻣﻦ
أﻣﻨﯿﺔ إﺑﺮاھﯿﻢ ﻣﺤﻤﺪ ﻋﻔﯿﻔﻲ ﻛﻠﯿﺔ طﺐ اﻟﻔﻢ واﻷﺳﻨﺎن ﺟﺎﻣﻌﺔ ٦أﻛﺘﻮﺑﺮ
ﻗﺴﻢ ﺑﺎﺛﻮﻟﻮﺟﯿﺎ اﻟﻔﻢ ﻛﻠﯿﺔ طﺐ اﻟﻔﻢ واﻷﺳﻨﺎن ﺟﺎﻣﻌﺔ اﻟﻘﺎھﺮة ٢٠١٢
Immunohistochemical Assessment of Cancer Stem Cells in Mucoepidermoid Carcinoma
Thesis Submitted to the Faculty of Oral and Dental Medicine, Cairo University in Partial Fulfillment of the Requirements for the Degree of Master in Basic Dental Science (Oral Pathology)
By
Omnia Ibrahim Mohammed Afifi B.D.S Faculty of Oral and Dental Medicine, October 6 University.
Department of Oral Pathology Faculty of Oral and Dental Medicine, Cairo University.
2012
Supervisors
Prof. Dr. Samia Mostafa El-Azab Professor of Oral Pathology Faculty of Oral and Dental Medicine, Cairo University
Dr. Rehab Fawzi Kasem Lecturer of Oral Pathology Faculty of Oral and Dental Medicine, Cairo University
ﺗﺤﺖ إﺷﺮاف اﻷﺳﺘﺎذ اﻟﺪﻛﺘﻮرة /ﺳﺎﻣﯿﺔ ﻣﺼﻄﻔﻰ اﻟﻌﺰب أﺳﺘﺎذ ﺑﺎﺛﻮﻟﻮﺟﯿﺎ اﻟﻔﻢ ﻛﻠﯿﺔ طﺐ اﻟﻔﻢ واﻷﺳﻨﺎن ﺟﺎﻣﻌﺔ اﻟﻘﺎھﺮة
اﻟﺪﻛﺘﻮرة /رﺣﺎب ﻓﻮزي ﻗﺎﺳﻢ ﻣﺪرس ﺑﺎﺛﻮﻟﻮﺟﯿﺎ اﻟﻔﻢ ﻛﻠﯿﺔ طﺐ اﻟﻔﻢ واﻷﺳﻨﺎن ﺟﺎﻣﻌﺔ اﻟﻘﺎھﺮة
First of all, thanks to Allah for guiding and helping me during this work. There is no word capable of expressing my gratitude and appreciation to Prof. Dr. Samia Mostafa Al-Azab, Professor of Oral Pathology, Faculty of Oral and Dental Medicine, Cairo University, for resolving the most difficult obstacles I met throughout this work. I shall never forget her wise guidance, continuous encouragement, close supervision and most valuable advices. I would like to express my sincere gratitude to my supervisor Dr. Rehab Fawzi Kasem, Lecturer of Oral Pathology, Faculty of Oral and Dental Medicine, Cairo University, for her valuable instructions,
endless
patience
that
made
her
meticulously revise this work word by word and assistance in various processes of putting this study in its final form.
Omnia Ibrahim 2012
To my lovely parents who are the most beautiful thing in my life and whom I cannot find adequate words to express my gratitude and never be able to repay them for their patience, kindness and their great support toward me.
To my dearest brothers who without their support and continuous help, none of this work would have been accomplished.
I.
Contents
Chapters
Page
Introduction and Review of literature
1
Stem Cells.
1
Types of Stem Cells.
3
Stem Cell Niche.
13
Identification of Stem Cells.
14
Cancer Stem Cells.
16
Cancer Stem Cells Hypothesis.
17
Origin of Cancer Stem Cells.
21
Identification of Cancer Stem Cells.
25
Expression of Stem Cell Markers in Different Human Tumors.
27
CD133 as a CSCs Marker.
30
Expression of CD133 as a Stem Cell Marker in Different Human Tumors. Expression of Other Stem Cell Markers in Different Salivary Gland Tumors.
32
Aim of the Study
35
39
I. Contents (continue) Chapters
Page
Materials and Methods
40
Results
45
Discussion
63
Summary 69
Conclusion
71
References
72
II.
List of Tables
Numbers
Contents
Page
CSC Markers and their
26
of tables
(1)
sources.
(2)
Statistical
difference
in
CD133+ colonies of cells
48
between low and high grade MEC.
(3)
Individual scoring data for the samples of low grade MEC
55
that represent CD133+ cells.
(4)
Individual scoring data for the samples of high grade MEC that represent CD133+ cells.
49
III.
Numbers
List of Graphs
Contents
Page
Scores of CD133+ colonies of cells in
50
of graphs
(1)
low and high grade MEC.
IV.
List of Figures
Numbe rs
Figure caption
of
Pa ge
figures 1
Stem Cell replication and differentiation.
1
2
Stem cells properities.
1
3
Stem Cells Potency.
3
4
Characteristics of ES cells.
4
5
HSCs.
6
6
UCB Stem Cells.
7
7
MSC’s are capable of differentiating into mesenchymal tissues.
8
8
DSCs.
9
9
NSCs
10
10
iPSC.
12
11
Tumor cells are heterogenous and only CSCs can proliferate extensively and form new tumors.
19
IV. List of Figures (continue) Numbe rs
Pa
Figure caption
of
ge
figures 12
Stem cell specific and conventional cancer therapies.
20
13
Origin of CSCs
24
14
A photograph illustrating the structure model of CD133.
31 50
15 Photomicrograph
of
low
grade
MEC
(H&E×100). 50
16 Photomicrograph
of
low
grade
MEC
(H&E×100).
51
17 Photomicrograph (H&E×100).
of
low
grade
MEC
51
18 Another field of previous photomicrograph of low grade (H&E×200).
52
19 Photomicrograph
of
low
grade
MEC
(H&E×200). 52
20 Photomicrograph of low grade MEC (H&E×200).
53
21 Another case of low grade MEC (H&E×100).
IV. List of Figures (continue) Numbe rs
Pa
Figure caption
of
ge
figures 53
22 Higher
magnification
photomicrograph
of
of low
grade
previous MEC
(H&E×200). 54
23 Another photomicrograph of low grade MEC (H&E×400).
56
24 Photomicrograph
of
high
grade
MEC
(H&E×100). 56
25 Photomicrograph
of
high
grade
MEC
(H&E×200). 57
26 Photomicrograph of another case of high grade MEC (H&E×200).
57
27 Photomicrograph
of
high
grade
MEC
(H&E×400). 58
28 Another case of high grade MEC (H&E×400).
58
29 Another field of the previous case (H&E×400).
59
30 Photomicrograph of low grade MEC (antiCD133×200).
59
31 Photomicrograph of another case of low grade MEC (anti-CD133×200).
IV. List of Figures (continue) Numbe rs
Figure caption
of
Pa ge
figures 61
32 Photomicrograph of low grade MEC (antiCD133×400).
61
33 Photomicrograph of high grade MEC (antiCD133×200).
62
34 Photomicrograph of high grade MEC (antiCD133×200).
62
35 Photomicrograph of high grade MEC (antiCD133×200).
V. List of Abbreviations Abbreviations
Words
ABCG2
ATP-binding cassette superfamily G member 2
Abs
Antibodies
ACC
Adenoid cystic carcinoma
AECs
Amniotic stem cells
ALDH
Aldehyde dehydrogenase
AML
Acute Myeloid Leukemia
ASCs
Adult stem cells
ATP
Adenosine triphosphate
B-SA
Biotin Streptavidin Amplified Detection System
BM
Bone Marrow
Bmi-1
Polycomb ring finger-1
Ca
Calcium
CD
Cluster of differentiation (cluster of designation)
CK5/14
Cytokeratin 5/14
CML
Chronic Myeloid Leukemia
CNS
Central Nervous System
CRC
Colorectal cancer
CSCs
Cancer Stem Cells
DNA
DeoxyriboNucleic Acid
DSCs
Dental stem cells
ESA
Epithelial specific antigen
ES cell line
Embryonic Stem Cell Lines
ESCs
Epidermal Stem Cells
FACS
Fluorescence Activated Cell Sorting
FSCs
Fetal Stem Cells
GIT
Gastro-intestinal tract
G phase
Growth phase
H&E
Hematoxylin and Eosin
HIF-1α
Hypoxia-inducible factors-1α
HMGA2
High-mobility group AT-hook 2 gene
HNSCC
Head and Neck Squamous Cell Carcinoma
HSCs
Hematopoietic Stem Cells
IHC
Immunohistochemistry
kDa
kilodalton
LGR5
Leucine-rich repeat-containing G-protien coupled Receptor 5
MB
Medulloblastoma
MEC
Mucoepidermoid Carcinomas
M phase
Mitosis phase
V. List of Abbreviations (continue) Abbreviations
Words
MSCs
Mesenchymal Stem Cells
NCAM
Neural Cell Adhesion Molecule
NCSCs
Neural Crest Stem Cells
NGF
Nerve growth factor
NK-cells
Natural Killer Cells
NOD/SCID
Non-obese diabetic /Severe Combined Immunodeficiency
NSCs
Neural Stem Cells
Oct-4
Octamer binding protien-4
OSCCs
Oral Squamous Cell Carcinoma
PBS
Phosphate buffered saline
PCR
Quantitative Reverse-Transcriptase
pH
Potential Hydrogen (a measure of acidity and alkalinity)
PSA-NCAM
Polysialic acid-neural cell adhesion molecule
P53
Protein 53
p75 NTR
p75 Neurotrophin R
Q RT-PCR
Quantitative Real-Time Polymerase Chain Reaction
Sca-1
Stem cell antigen-1
