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Prostate CancerProstate Cancer
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Graduation research Project
Partial Fulfillment for the degree of B.Sc. In
Ecology/Biochemistry
Prepared By
Ahmad Salah Roushdy
Supervised By
Professor Ahmad R. Bassiouny
May 2006
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What is the Prostate?What is the Prostate?
A male sex gland
The size of a walnut below the bladder and in
front of the rectum
Produces the fluid that is
part of semen
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Risk Factors for Prostate CancerRisk Factors
for Prostate Cancer
Age Found mainly in
men over age 55. Average
age of diagnosis is 70
Family History Mens
risk is higher if father orbrother is diagnosed
before the age of 60
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Risk Factors continuedRisk Factors continued
Race Prostate cancer is found more often in
African American men then White men. It is less
common in Asian and American Indian men
Dietary factors Evidence suggests that a diet
high in fat may increase the risk of prostate cancerand diets high in fruits and vegetables decrease the
risk
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Symptoms of Prostate Cancer
Symptoms of Prostate Cancer
1. Frequent urination
2. Inability to urinate3. Trouble starting and stopping urination
4. Blood in the urine or semen
5. Painful ejaculation
6. Painful or burning urination
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What Goes WrongWhat Goes Wrong
1. Prostate infections prostatitis
Fairly common in men from the teen years on.
These infections can be brief or long-lasting, mild or severe,
easy or difficult to treat with antibiotics.
Symptoms ofprostatitis can include frequent and/or painful
urination, other urinary difficulties, or pain during sex.
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What Goes Wrong continuedWhat Goes Wrong continued
2. Prostate enlargement benign prostatic hyperplasia.
Unwanted but non-cancerous enlargement of the prostate.
Although men in their twenties can suffer from BPH, it
usually surfaces later in life.
It's estimated that half of all men have BPH by the age of
60, and 90% will suffer from it by age 85
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What Goes Wrong continuedWhat Goes Wrong continued
3. Prostate cancer
Prostate cancer is a malignant tumor that usually
begins in the outer part of the prostate. In most
men, the cancer grows very slowly.Urinary
Bladder
Ejaculatory
Duct
Urethra
Transition
Zone
(BPH)
Central
Zone
Anterior
Fibromuscular
Stroma
Peripheral Zone
(Prostate Cancer)
Rectum
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Detecting Prostate Cancer
Prostate cancer rarely causes symptoms early in the
course of the disease.
Patient may present with advanced and metastatic
prostate cancer.
Today, the majority of prostate cancers are detected
based on Digital Rectal Examination DRE
abnormalities or Prostate Specific Antigen PSA
elevations.
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Normal Function of PSA
Secreted in high concentrations into the seminal fluid
(mg/mL), where it cleaves gel-forming proteins
semenogelin andfibronectin to increase sperm
motility
Normally found in low concentration in sera (ng/mL).
Serum PSA increases as epithelial cell number (tumor
volume) increases.
Pro-PSA (Inactive)
PSA (Active)
Inhibitory Region
Pro-Seminogelin (Inactive) Seminogelin (Active)
Liquefaction of Seminal Fluid
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Forms of PSA in Blood
70-90% of serum PSA is boundto the protease
inhibitor1-Anti-Chymotrypsin (ACT).
10-30% of serum PSA is free and unbound.
Minor amounts (
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Use of PSA as a Marker for
Prostate Cell Growth
PSA elevations may indicate the presence of prostate
disease.
Normal prostatic growth: 0.04ng/mL increase peryear.
BPH: 0.07-0.27ng/mL increase per year.
Prostate Cancer: greater than 0.75ng/mL increase
per year.
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normalnormal
prostateprostate
epitheliumepithelium
histologic
prostate
cancer
localized
prostate
cancer
Progression of Prostate Cancer
tumor suppressor geneinactivation / mutation?
androgen
independent
cancer
metastatic
prostate
cancer
curable incurable
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Genetic Epidemiology ofGenetic Epidemiology of
Prostate CancerProstate Cancer is prostate cancer familial? ..... yesis prostate cancer familial? ..... yes
is the familial clustering compatible withis the familial clustering compatible withMendelian inheritance? ..... yesMendelian inheritance? ..... yes are there susceptibility genes responsible for thisare there susceptibility genes responsible for this
pattern of inheritance? ....?pattern of inheritance? ....?
Use linkage analysis to identify and mapUse linkage analysis to identify and map what can identification of these genes tell us aboutwhat can identification of these genes tell us about
the causes of prostate cancer?the causes of prostate cancer?
Inflammation??Inflammation??
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Novel Prostate CancerNovel Prostate Cancer
GenesGenes
RNASEL (chr1)RNASEL (chr1) (Carpten et al 2002)(Carpten et al 2002)Interferon inducible anti viral defenseInterferon inducible anti viral defense
MSR1MSR1 (chr 8) (Xu et al 2003)(chr 8) (Xu et al 2003)Binds diverse ligands including gram -/+ bacteriaBinds diverse ligands including gram -/+ bacteria
Both of these genes are part of the innate immuneBoth of these genes are part of the innate immuneresponse pathway to fight infectionresponse pathway to fight infection
Mutations inMutations inMSR1MSR1 andandRNASELRNASEL are responsibleare responsiblefor ~10% - 15% of hereditary prostate cancer infor ~10% - 15% of hereditary prostate cancer inJHU family collectionJHU family collection
Mixed results seen in other study populationsMixed results seen in other study populations
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Early Detection ofEarly Detection of
Cancer, Does it makeCancer, Does it make
any difference?any difference? Despite recent progress in treatment, Cancer is theDespite recent progress in treatment, Cancer is the
second leading cause of mortality in developedsecond leading cause of mortality in developed
countries.countries.
Early Detection of Cancer results in better treatmentEarly Detection of Cancer results in better treatment
options and higher survival rates.options and higher survival rates.
Unfortunately, current Cancer diagnostics oftenUnfortunately, current Cancer diagnostics often
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Cancer BiomarkersCancer Biomarkers
They are important tools for Cancer detectingThey are important tools for Cancer detecting
and monitoring.and monitoring.
They serve as hallmarks for the physiologicalThey serve as hallmarks for the physiological
status of a cell at a given time and changestatus of a cell at a given time and change
during the disease progression.during the disease progression.
Used routinely for population screening,Used routinely for population screening,
disease diagnosis, prognosisdisease diagnosis, prognosis..
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Some Established Cancer BiomarkersSome Established Cancer Biomarkers
Hepatoma; testicularcancer
Colon; breast; lung;pancreatic
Prostate
Ovarian
BreastGastrointestinal
Testicular cancer
Alpha-fetoprotein AFP)
Carcinoembryonic antigen(CEA)
Prostate-specific antigen(PSA)
CA125
CA15.3
CA19.9Chroriogonadotropin (hCG)
Cancer typeBiomarker
But none of them has adequate sensitivity, specificity
and predictive value for population screening which leads to
many false-positive results.
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Over many years of developing Cancer Biomarkers, It wasOver many years of developing Cancer Biomarkers, It was
postulated that a molecule may become a reliable Cancerpostulated that a molecule may become a reliable Cancer
Biomarker if it has certain characteristicsBiomarker if it has certain characteristics
1. It should be a secreted or shed protein.
2. Able to diffuse into the circulation during tumor
development and progression.
3. Such proteins should be stable (not degradable).
4. Shouldnt be bound to any inhibitor which could
interfere with their measurements.
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? Why Cancer Proteomics? Why Cancer Proteomics
DNA tells what possibly,DNA tells what possibly,
RNA what probably andRNA what probably and
Proteins what actually happens.Proteins what actually happens.
DNA sequence does not predict if the protein is in an active form.
RNA quantitation does not always reflect corresponding protein levels.
Multiple proteins can be obtained from each gene (alternative
splicing).
Genomics cannot predict post-translational modifications and the
effects thereof.
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Surface-enhancedSurface-enhanced
laser desorption/ionization-laser desorption/ionization-time of flight-mass spectrometry.time of flight-mass spectrometry.
SELDI ProteinChip is a mass spectrometrictechnology that accomplishes protein separation
on a chromatographic chip surface by binding
subsets of proteins from complex mixtures.
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SELDI-TOF-MS consists of 3 majorSELDI-TOF-MS consists of 3 major
componentscomponents
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The array contains either 8 or 16 spots
comprised of a specific
chromatographic treated surface.
Chemically-treated surface
Binds
hydrophobic
molecules
Binds +ve charged
proteins; rich in
Lys, Arg, His
Binds -ve charged
proteins; rich in
Asp, Glu
Proteins that bind
to these metal ions
will be bound to
the chip
General protein
binding surface
which bind via
Ser, Ther, Lys
Chemically-treated surface
Binds
hydrophobic
molecules
Binds +ve
charged proteins;
rich in Lys, Arg,
His
Binds -ve charged
proteins; rich in
Asp, Glu
Proteins that bind
to these metal
ions will be
bound to the chip
General protein
binding surface
which bind via
Ser, Ther, Lys
(Antibody - Antigen) (Receptor - Ligand) (DNA - Protein)
Biologically treated surface
(Antibody - Antigen)(Receptor - Ligand) (DNA - Protein)
Biologically treated surface
Chemically treated surfaces will retain whole classes of
proteins, while Biologically treated surfaces are designed
to interact specifically with a single target proteins
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The array is then inserted into the ProteinChip Reader
The ProteinChip Reader is a Laser Desorption/Ionization TOF - MS
Pulsed UV
Nitrogen
Laser source
When
laser is
activated
The sample becomes
irradiated, desorption and
ionization proceeds toliberate aseous ions
These gaseous ions enter the TOF-
MS region, which measures the
mass/charge of each protein based
on its velocity through an ion
chamber
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A) Detected proteins are displayed as a series of peaks showing relative
abundance of the detected proteins versus their MWs.
Spectra View
B) To identify differences between the detectd prtoeins, a software has beendeveloped to generate a simulated 1D gel electrophoresis
Gel View
C) Based on the spectra view, a map view will be also generated for better
identification
Map View
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Protein mapping for a certain disease
By using different chemically surface-treated arrays for the same sample.
Each surface will retain different groups of proteins depending on their
physiochemical properties as well as the pH of the sample.
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The end-result of a SELDI-TOF-MS analysis is a list of the
MWs of the proteins whose relative abundance differ
significantly between 2 or more samples.
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The identity of the peak masses used in the tree analysis pattern is
not necessary for making a diagnosis.
The only requirement for this classification system to make an
accurate diagnosis is that the biomarkers be reproducibly detected by
SELDI and accurately selected by the classifier.
However, because knowing their exact identities will be essential
for understanding what biological roles of these peptide/proteins may
have in the cancer, efforts are under way to purify, identify, and
characterize these protein/peptide biomarkers.
(European Urology 47 (2005) 456462)
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The ability to simultaneously test forThe ability to simultaneously test for
multiple protein changes by themultiple protein changes by the
ProteinChip SELDI system increases theProteinChip SELDI system increases the
diagnostic sensitivity, and withdiagnostic sensitivity, and with
Biomarker Pattern Software, has theBiomarker Pattern Software, has the
potential to improve the Early diagnosispotential to improve the Early diagnosis
of Prostate Cancer.of Prostate Cancer.
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