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Tumor Markers
Tumor Markers
Lecturer:
Chatchawin PETCHLERT, Ph.D.
Head, Department of Biochemistry
Deputy Head, Department of Biotechnology
Department of BiochemistryFaculty of Science, Burapha University
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Department of BiochemistryFaculty of Science, Burapha University
Tumor Markers
DNA structure andorganization within the cell.
The complement of 46
metaphase chromosomes
(human karyotype) is shown
lying next to an intact
nucleus. One chromosome is
blown up to reveal the degreeof chromatin condensation
and associated nucleosome
structures involved in the
organization of eukaryotic
DNA. The red box shows
the molecular structure of the
nucleoside bases which formthe struts of the DNA helix,
and the bonds between the
pyrimidine and purine bases(dashed lines).
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Department of BiochemistryFaculty of Science, Burapha University
Tumor Markers
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Department of BiochemistryFaculty of Science, Burapha University
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Department of BiochemistryFaculty of Science, Burapha University
Tumor Markers
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Department of BiochemistryFaculty of Science, Burapha University
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Department of BiochemistryFaculty of Science, Burapha University
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A tumor is an abnormal growth of body tissue. It
originally meant any abnormal swelling, lump or
mass.
Tumors can be cancerous (malignant) or non-
cancerous (benign).
Tumor has become synonymous with neoplasm
except some neoplasms, such as leukemia do not
form tumors.
Tumor
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Faculty of Science, Burapha UniversityTumor Markers
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Faculty of Science, Burapha UniversityTumor Markers
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Faculty of Science, Burapha UniversityTumor Markers
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The term cancercancer derives from the Latin forcrabdue to the resemblance of the swollen veins around atumour to a crabs limbs.
Cancer is not a single disease; there are over 200 pathologically
distinct neoplastic entities (each depending on the cell/tissue-type
of origin). Nevertheless, all cancers are generally characterized byuncontrolled abnormal cell growth, which eventually forms a cell
mass, or tumour. Tumour development, or tumourigenesis,
generally occurs over a period of years, and cancer incidence is
more prevalent in aged populations. Sooner or later, the tumour
will generally invade and destroy surrounding normal tissues, andcells from the tumour can spread (metastasize) through the
bloodstream or lymphatic system to start new tumours in otherparts of the body.
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Liver cancer Colon cancer Gastric cancer
Skin cancer Breast cancerCervical cancer
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Faculty of Science, Burapha UniversityTumor Markers
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The process of carcinogenesis,that is, the change in cells from normal,
controlled cell division and differentiation
to cells that are transformed, dividing
without check, and are undifferentiated or
abnormally differentiated, does not
appear to occur as a single step. In other
words, transformation is a multistageprocess and involves a sequence of
events from tumour cell initiation, to
promotion, malignant conversion and
progression. Evidence for this comes
from in vitro studies (e.g. cell
transformation studies), animal models
and clinical/epidemiological observationsand, in particular, the long latency period
between cell initiation and the
appearance of a tumour in the target
tissue and the progression to metastaticdisease
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ProtoProto--oncogeneoncogene can become activated to anoncogene by a relatively small modication of itsoriginal function.
There are three basic activation types:
Mutation within a proto-oncogene can cause a
change in the protein structure, causing
an increase in protein (enzyme) activity and/or
loss of gene regulation.
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An increase in protein activity, caused by an increase in gene expression
an increase of protein stability, prolonging its existence and thus its
activity in the cell
a gene duplication resulting in an increased amount of protein in the cell.
A chromosomal translocation causing
inappropriate or constitutive gene expression, that is, in the wrong cell
type or at the wrong time; for example, c-Abl in chronic myeloid leukemia
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Tumour suppressor genesTumour suppressor genes encode for proteins whichgenerally inhibit tumorigenesis, and their existence was
originally inferred from experiments where tumour cells were
fused with normal cells in culture that resulted in non-
tumourigenic hybrids.
Subsequent loss of specic individual chromosomes or
specic regions of a chromosome in the hybrid, and derived
from the normal parental cell, resulted in the re-emergence
of the tumorigenic phenotype.
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Numerous tumour suppressor genes have since beenidentied, and the functions of the associated proteinsfall into several categories:
1. The activities of the cyclin-dependent kinase (CDK) family are in turn regulated by
cyclin-dependent kinase inhibitors (CDIs), which are essential for continuing the cell cycle. Ifthese genes are not expressed the cell cycle will arrest, effectively inhibiting cell division (Fero
et al., 1998).
2. Cell cycle regulation and DNA damage surveillance: the cell cycle can be arrested by
tumour suppressor genes of the p53 and Rb pathways that are activated by aberrant
proliferative stimuli or DNA damage (reviewed by Kohn, 1999; Mirza et al., 2003). After cell
cycle arrest, there is an opportunity to rectify DNA damage where necessary. The role of somemembers of the p53 and Rb pathways is to maintain low levels of inactive TP53 in the absenceof genomic damage allowing normal cell cycle function.
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3. If the DNA damage is irreparable, the cell enters senescence or undergoes apoptosis(programmed cell death), preventing replication of the damaged genome (Malumbres and
Barbacid, 2001). Apoptosis in multicellular organisms involves a series of biochemical
events leading to a characteristic cell morphology and death, characterized by a variety of
morphological changes, including blebbing, changes to the cell membrane such as loss of
membrane asymmetry and attachment, cell shrinkage, nuclear fragmentation, chromatin
condensation and chromosomal DNA fragmentation. Mutations in genes regulating
apoptosis can shift the balance between pro- and anti-apoptotic inuences, and maycontribute to genomic instability, if cells carrying DNA damage are permitted to survive.
Such cells which survive past their use-by-date are capable of replication and they and
their progeny are subject to increased genomic instability, increasing the likelihood of the
cell becoming cancerous or diseased (Schmitt et al., 2007).
4. Some tumour suppressor genes encode for proteins involved in cell adhesion which
prevent tumour cells from dispersing, block loss of contact inhibition, and inhibit metastasis.These proteins are known as metastasis suppressors.
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The scientific term to describe an abnormalproliferation of genetically altered cells.
Neoplasms can be benign or malignant:1) Malignant neoplasmormalignant tumor:synonymous with cancer.
2) Benignneoplasm or benigntumor :atumor (solid neoplasm)that stops growing byitself, does not invade other tissues and doesnot form metastases.
Neoplasm
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Tumor markers are molecules occurring in blood,
urine or body tissue that are associated with
cancer.
A tumormarkermaybemadeby a tumoritself or
by the body in response to the tumor.
Tumor marker measurement or identification isuseful in patient diagnosis or clinical management.
Tumor markers
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Tumor markers can be classified in two groups:
1) Cancer-specific markers
2) Tissue-specific markers
Classification
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Cancer-specific markers arerelatedto the
presence of certaincanceroustissue.
Becausethere is a largeoverlapbetween themanydifferenttumortissuetypes and the
markersproducedthesemarkersmightnotbe
specific in making a diagnosis.
Cancer-specific markers
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Tissue-specific markers are related to specific
tissues which have developed cancer.
These substances are not specifically related tothe tumor, and may be present at elevated levels
when no cancer is present.
Tissue-specific markers
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Tumor marker tests are not used alone to
detect and diagnose cancer because
: most tumor markers can be elevated
in patients who don't have a tumor;
: no tumor marker is entirely specificto a particular type of cancer;
: not every cancer patient has an
elevated tumor marker level, especiallyin the early stages of cancer, when
tumor marker levels are usually stillnormal.
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Tumor markers can be used for one of four
purposes:
(1) screening a healthy population or a high riskpopulation for the presence of cancer;
(2) making a diagnosis of cancer or of a specifictype of cancer;
(3) determining the prognosis in a patient;
(4) monitoring the course in a patient in remission
or while receiving surgery, radiation, orchemotherapy.
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Department of Biochemistry
Faculty of Science, Burapha UniversityTumor Markers
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Types ofTumor Markers
1. Enzymes and isoenzymes
2. Hormones
3. Immunoglobulins4. Antigens (oncofetal and CHO)
5. Receptors
6. Oncogene products
7. Genetic markers
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CEA is a protein found in manytypes of cells but
associated with tumors and the developingfetus
(oncofetal antigen).
It is a complex glycoprotein of molecular weight~150-300 kDa, that is associated with the plasma
membrane of tumor cells, from which it may be
released into the blood.
CEA is most useful as a marker for colorectal,
pancreatic, gastric, lung, breast, and ovarian
carcinoma.
Carcinoembryonic antigen(CEA)
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AFP is a normal fetal serum protein synthesized by the
liver, yolk sac, and gastrointestinal tract that sharessequence homology with albumin .
The level of AFP is typicallyhigh in the fetus's blood. It
goesdown in the baby's blood afterbirth. And by a year
of age, it is virtuallyundetectable.
It is a glycoprotein with molecular mass of70 kDa.
AFP is elevated in normal pregnancy, benign liverdisease (hepatitis, cirrhosis), as well as in cancer.
AFP is of importance in diagnosinghepatocellular
carcinoma and maybeuseful in screeningprocedures.
Alpha-Fetoprotein (AFP)
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PSA is prostate-specific, notcancer-specific.
It is found in normal prostatic epithelium and secretionsbut not in other tissues. It is a glycoprotein.
A variety of conditionscanraisePSAlevels:prostatitis
(prostate inflammation), benign prostatic hypertrophy
(prostateenlargement), and prostate cancer.
PSA is highlysensitive for the pesence of prostaticcancer.
Prostate-SpecificAntigen(PSA)
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HCG is a glycoprotein consisting of subunits a- and b-subunits, which are non-convalently linked.
The hormone is normally produced by the
syncytiotrophoblastic cells of the normal placenta and is
elevated in pregnancy.
Its most important uses as a tumor marker in trophoblastictumor(tumors of placenta) and some tumors of the testis.
The level of HCG is occasionally elevated in other cancersincluding those of breast, lung, and gastrointestinal tract.
Human chorionic gonadotropin (HCG)
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CA 19-9is synthesized by normal human pancreatic
and biliary ductular cells and by gastric, colon, and
salivary epithelia.
This carbohydrate antigen is a glycolipid with amolecular mass > 200 kDa.
CA 19-9 is a marker for both colorectal, pancreatic
carcinoma and cholangiocarcinoma.
Carbohydrate antigen CA 19-9
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CA1
25 is high molecular mass (> 200
kDa)glycoprotein.
CA 125 is most useful as a marker for ovarian
cancer.
CA 125 is often elevated in patients with ovarian
cancer, its level following the patient's clinical
course. With surgical resection or chemotherapy,the level correlates with patient response.
The CA 125 is elevated in other cancers including
endometrial, pancreatic, lung, breast, and coloncancer, and in menstruation, pregnancy.
Carbohydrate antigen CA 125
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Neuron specific enolase is an isozyme of the
glycolytic pathway that is found only in brain andneuroendocrine tissue.
NSE is found in tumors associated with aneuroendocrine origin, including small lung cancer,neuroblastoma and etc.
Use of NSE has been evaluated in lung cancer andneuroblastoma.
NeuronSpecificEnolase(NSE)
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Department of Biochemistry
Faculty of Science, Burapha UniversityTumor Markers
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Samples used for tumor marker estimations
1- Blood, plasma, and serum
2- Pleural fluid, ascetic fluid, or
pericardial fluid
3- Urine
4- CSF
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Gene Expression
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Control of gene expression
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Why Proteins ?
Genome (all genes): What couldhappen
Transcriptome (all mRNAs):
What might be happening
Proteome (all proteins):
What is happening
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What is Proteomics ?
A newly emerging field of life
science research that uses high
throughput (HT) technologies todisplay, identify and/or characterize
all the proteins in a given cell, tissue
or organism.
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Content
Proteomics (2D-PAGEMS) Cell lysis and Protein
extraction / Prefractionationand enrichment
Protein separation/Proteindetection
Image analysis
Protein identification
Proteomics and itsapplications
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2D-Polyacrylamide Gel
Electrophoresis (2D-P
AGE)
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2D-Polyacrylamide Gel
Electrophoresis (2D-P
AGE)-+
pH 3.0 10.06.5 103.0 5.5 8.5
100
75
50
37
20
10
150
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Protein
separation
Protein
staining
sample
preparation
Image analysis Protein identification54
2D-Polyacrylamide Gel
Electrophoresis (2D-P
AGE)
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Patient Healthy
Proteomic of CA patient and healthy
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Biomarker discovery
Rheumatoidarthritis Infectiousmeningitis
Prostatedisease Ovarian cancer
Pancreatic cancer Systemiclupuserythematosus
Leukemia Cervical cancer Alzheimersdisease Denguevirusinfection
Hepatocellularcarcinoma HIVinfection
Lung cancer WestNilevirus
Melanoma RenalcellcarcinomaHypertension Breast cancer
Raghothama Chaerkadyand Akhilesh Pandey. Annu Rev Path Mech Dis 2008
3
: 485
-498
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Protein biomarker
Righetti et al., 2005Clin Chim Acta 357 : 123-139 58
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Metabolic Pathways
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Detection of tumor markers by ELISA
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