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INTRODUCTION
CANCER
Cancer is an overgrowth of cells bearing cumulative genetic injuries that confer growth advantage over the normal cells [Nowells Law] Cancer cells can be characterized as antisocial, fairly autonomous units that appear to be indifferent to the constraints and regulatory signals imposed on normal cells [Robbins]
NORMAL CELLS
Blood vessel
Loss of contact inhibition Increase in growth factor secretion Increase in oncogene expression Loss of tumor suppressor genes
Oncogene expression is rare Intermittent or co-ordinated cogrowth factor secretion Presence of tumor suppressor genes
CHARACTERISTICS OF CANCER
Clonality Autonomy Anaplasia Metastasis
Cancer Genetics
Tumors arise as clones from a single cell. At the cellular level, cancer is a genetic disease. The development of the malignant clone is due to mutations in DNA due to:
Random replication errors Exposure to carcinogens Faulty DNA repair process
CHARACTERISTICS OF CANCER
Autonomy
Cancer cells are able to proliferate despite regulatory influences. Unrestricted proliferation results in tumor formation. Mechanisms:
Growth factor secretion Increased number of cell receptors Independent activation of key biochemical process
AUTONOMY
Brought about by mutations in the cells genetic apparatus Most common in tissues with rapid turnover, especially:
- those exposed to environmental agents - those whose proliferation is hormonedependent
G0
G2/M checkpoint
Mitosis
M
DNA content = 4n
DNA content = 2n
G2 S
DNA synthesis
G1
G1/S checkpoint
G2/M Checkpoint
Regulated by the cyclin B/cdc2 (mitosis promoting factor or MPF). Activity of this cyclin with its substrate results in:
Chromosome condensation Nuclear membrane breakdown Spindle formation
G1/S Checkpoint
Area most often disrupted in cancer. Mechanism of regulation is complex and involves the phosphorylation of the Rb gene. This results in:
Activation of several genes needed for S phase progression. Promotes differentiation through association with transcription factors.
Rb Gene Activation
Cyclin Regulators
Regulated by cdk inhibitors (cdki). May be induced by growth inhibitors and inhibited by positive growth factors. Genetic alterations in cdki occur with high frequency in some cancers.
Cyclin Regulators
p 21: inhibits cell cycle progression and permits DNA repair to take place. P53: the guardian of the genome
In the presence of DNA damage, influences transcription to either:
Halt cell cycle progression to facilitate DNA repair. In cases of severe DNA damage, activates apoptosis.
Mutations in p53 are the most common genetic alterations found in human cancer.
Angiogenesis
Process of new blood vessel formation. Clinical importance:
Tumor vessel number correlates positively with risk and degree of dissemination. Several cytokines that stimulate endothelial cell proliferation also stimulate proliferation of malignant cells.
Triad of Invasion
Adhesion with the basement membrane Local proteolysis Mobility and ability to translocate through dents in bodys structural barriers
MOLECULAR CARCINOGENESIS
Mutation
Gene Families in Cancer Development 1 - Oncogenes 2 - Tumor Suppressor genes 3 - Mutator genes
Cancer Genes
Proto-oncogenes normally promote normal cell growth; mutations convert them to oncogenes. Tumor suppressor genes normally restrain cell growth; loss of function results in unregulated growth. Mutator or DNA repair genes when faulty, result in an accumulated rate of mutations.
ONCOGENE FAMILY
+ oncogenes
dominant & highly conserved types: viral oncogenes [v-oncs] cellular oncogenes [c-oncs] Proto-oncogene Mutation Oncogene
ONCOGENE FAMILY
Classification of Oncogenes A. Secreted Growth Factors
c-sis, hst
C. Intracellular Transducers
c-src, c-abl, mst, ras
SIGNAL TRANSDUCTION
ONCOGENE FAMILY
Mechanisms of Oncogene Activation 1. Point Mutation
H-ras Normal Bladder ca [codon 12] CGC p Gly CTC p Val
H-ras GTP
2. Gene Amplification
Double minutes HSRs
Homogenously Staining regions
Normal copy
Multiple copies
ONCOGENE FAMILY
Mechanisms of Oncogene Activation 3. Gene Translocation Ex. Burkitts Lymphoma
ONCOGENE FAMILY
Mechanisms of Oncogene Activation 3. Gene Translocation Ex. Chronic Myelogenous Leukemia [CML]
ONCOGENE FAMILY
Mechanisms of Oncogene Activation 4. Viral Gene Integration
promoter
Viral promoter
inhibit growth and multiplication of mutated cells prevent neoplastic transformation recessive & highly conserved
pRb
105-KDa nuclear protein inhibits E2F [prevents G1 p S transition] inhibited by: phosphorylation viral oncoproteins [E1A, HPV E7]
location: 17p13.1 product: p53 protein [53 KDa] function: induces DNA repair or apoptosis mutation: point mutation > deletion results to: loss of function & extended lifespan of p53 Clinical conditions: carcinomas, Li Fraumeni Syndrome p53 inhibited by: E1B, HPV E6, mdm2
p53 in action
involved in ensuring the fidelity of replication function: checks for & corrects mismatched pairs mutation inefficient repair & replication leading increased propensity of oncogenes and tumor suppressor genes to undergo mutation first described in E coli [Mut-HSL system] Fischel, et al = Human homologs leads to the formation of Microsatellite Instability [MIN+]
In summary
..
MUTATOR GENES
ONCOGENES
TS GENES
CARCINOGENS
Occupation related causes Lifestyle related causes
Tobacco Diet Sexual practices
y y y y y y y y y y y y y y y y y y
X-rays Stress Toxins Sunlight Solvents Pollution Cigarette Pesticides Herbicides Medications Airline travel Radioactivity Food additives Polluted Foods High heat cooking Synthetic materials Household cleaners Environmental Chemicals Lots more
R.I.P
They only said it was dangerous. They didnt say it could be
lethal.
MENU
Site of Malignancy
Lung, skin, liver Mesothelium, lung Leukemia Bladder Lung Numerous locations Lung Lung, skin Angiosarcoma of liver
Mycotoxins
Liver Cancer
Cervical Cancer
Multifactorial Factors
Tobacco + Alcohol
Oral Cavity Cancer Esophageal Cancer
CARCINOGEN METABOLISM
Three Main Categories: I. Chemical Carcinogens II. Physical Carcinogens III. Viral Agents
Mutations
Cancer
CHEMICAL CARCINOGENESIS
Stages: Initiation - primary exposure
Promotion - transformation
Frank Cancer
normal cells are exposed to a carcinogen not enough to cause malignant transformation requires one round of cell division normal cells are exposed to a carcinogen
1. Direct-acting carcinogens 2. Indirect-acting carcinogens
procarcinogen
Cytochrome P450
Ultimate carcinogen
initiated cells are exposed to promoters promoters are not carcinogens ! properties of promoters
CHEMICAL CARCINOGENESIS
Direct-acting Carcinogens
~ ~ ~ ~
Procarcinogens
PAHs Aromatic amines & Azo dyes Aflatoxin B1 Nitrosamine & Amides Asbestos Vinyl chloride Chromium, nickel, other metals Arsenic
Promoters
Physical Carcinogenesis
Radiation-induced mutation in the host cell Transmits irreversible changes in gene expression to cell progeny
PHYSICAL CARCINOGENESIS
Ultraviolet Rays
UV-A = 320 - 400 nm UV-B = 280 - 320 nm UV-C = 200 - 280 nm
PHYSICAL CARCINOGENESIS
Ultraviolet Rays
UV-C filtered by ozone UV-B Inhibition of cell division inactivation of enzymes induction of mutations cell death at high doses Squamous cell cancer Basal cell cancer Melanocarcinoma
includes electromagnetic rays & particulate matter mechanism: o free radicals & mutations pathology: leukemias > thyroid ca > lung & breast ca resistant tissues: bone, skin and the GIT
PRE-IRRADIATION
POST-IRRADIATION
Viral Carcinogenesis
Viral carcinogens are classified into RNA and DNA viruses. Most RNA oncogenic viruses belong to the family of retroviruses that contain reverse transcriptase mediates transfer of viral RNA into virus specific DNA.
Viral Oncogenes
RETROVIRUS Oncogene Oncogene Viral RNA
REVERSE TRANSCRIPTASE
Viral DNA
TRANSCRIPTION
NUCLEUS
INSERTION
DNA
TRANSCRIPTION
Viral genome
Viral RNA
Oncogene
CELL MEMBRANE
CYTOPLASM
NEOPLASMS Cervical Ca, warts, anogenital carcinoma Cervical carcinoma NPCa, African Burkitts Kaposis sarcoma Hepatocellular Ca Certain B cell lymphomas
Human papilloma virus Herpes simplex virus II Epstein-Barr virus Herpes simplex virus 8 Hepatitis B virus Herpes simplex virus 6 (HBLV)
NEOPLASMS
Over-expression of E6 & E7
in Burkitts, B-cell & Hodgkins lymphomas + NP ca tropism: CD21+ cells [e.g., B cells, epithelial cells] mechanism: viral entry episomal existence latency (+) LMP-1, EBNA-1, EBNA-2 immortalization
induction of chronic hepatocyte injury (+) HBx HBx activates protein kinase c for transformation
SIGNAL TRANSDUCTION
PROLIFERATION (Growth Factor Signaling Pathway) Receptor Tyrosine kinase Pathway (RTK)-Main pathway RTK ligands: NGF PDGF FGF EGF Functions of RTK: 1. promotion of cell survival 2. regulation of cell proliferation and differentiation 3. modulation of cellular metabolism
PROLIFERATION (Growth Factor Signaling Pathway) RTK SIGNALING PATHWAYS Ras-MAP Kinase Pathway- most prominent PI3 kinase Pathway Phospholipase C Pathway
PROLIFERATION
G0
G2/M checkpoint
Mitosis
M
DNA content = 4n
DNA content = 2n
G2 S
DNA synthesis
G1
G1/S checkpoint
G2/M Checkpoint
Regulated by the cyclin B/cdc2 (mitosis promoting factor or MPF). Regulated mainly by intracellular signal (Completion of DNA Synthesis) MPF is activated by dephosphorylation by cdc25 Cyclin B is degraded by Anaphase Promoting Complex (APC) Role of G2/M checkpoint: to prevent mitosis when DNA is damaged and not yet repaired
G1/S Checkpoint
Area most often disrupted in cancer. Mechanism of regulation is complex and involves the phosphorylation of the Rb gene. Regulated by extracellular signals (e.g. GF) R point (restriction)- point late in G1 beyond which cell cycle progression becomes independent from external GF Regulated mainly by CDK4/cyclin D
Rb Gene Activation
Cyclin Regulators
p 21: activated by p53 inhibiting cell cycle progression and permitting DNA repair to take place. P53: the guardian of the genome
In the presence of DNA damage, influences transcription to either:
Halt cell cycle progression to facilitate DNA repair. In cases of severe DNA damage, activates apoptosis.
Mutations in p53 are the most common genetic alterations found in human cancer.
p53 in action
CELL-CYCLE PROGRESSION
Clinical Significance
Oncogenic alterations in cell cycle regulators: Loss of p53 and pRB function as tumor suppressors Increased expression of Cyclin D1(Mantle Cell Lymphoma) CDK4 amplification in sarcomas, glioma Mutations in p16-binding domain of CDK4(Familial Melanoma) Inactivation of INK4 Alterations in Cyclin D1,p16 Decreased levels of p27 (Breast Ca) Over expression of cdc25
Therapeutic Implications
Approaches using Inhibitors of CDKs as therapeutic agents Small molecules Protein therapy Antisense Gene therapy Most cytotoxic agents block the cell cycle in the S/G2/M phases
IMMORTALIZATION
APOPTOSIS
1. 2. APOPTOSIS programmed cell death Important in: Steady-state kinetics of normal tissues Focal deletion of cells during normal embryonic development 3. Seen after chemotherapy and radiation * Balance between proliferation and apoptosis is critical in determining growth or regression
APOPTOTIC PATHWAYS
1) FAS-mediated apoptosis FAS cell surface receptor of TNF family which binds to FAS-L Eliminates unwanted activated T cells Pathway for cytotoxic-mediated signaling 2) P53-mediated apoptosis important after chemotherapy and radiation Induction of BAX and downregulation of BCL-2 Induced expression of FAS and DR5
Clinical Significance
Over expression of BCL-2 as a prognostic indicator Mutations of BAX in GI Ca and leukemias P53 provides a link between cell proliferation and apoptosis Cell survival signals: NF BCL-2 P53 mutations confer chemoresistance
EVADING APOPTOSIS
Therapeutic Implications
Antisense oligonucleotide against BCL-2 in the treatment of lymphoma BCL-2 antisense as chemosensitizing agent in solid tumors TRAIL ( TNF-related apoptosis inducing ligand) to induce apoptosis
ANGIOGENESIS
Formation of new blood vessels from existing vascular bed Carried out by endothelial cells (EC) and extra cellular matrix (ECM) Regulated by angiogenic factors (inducers and inhibitors) * A tumor is unable to grow larger than 1 mm3 w/o developing a new blood supply
Components of Angiogenesis
1) ENDOTHELIAL CELLS Fenestrated Increased cell adhesion molecules ( E-selectin) Increased integrins 3 essential for viability during growth Activated ECs release: bFGF PDGF IGF-1
Components of Angiogenesis
2) INDUCERS OF ANGIOGENESIS
VEGF main inducer TGF TNF- low concentration - inducer high concentration inhibitor PDGF/thymidine phosphorylase TGF EGF IL-8
Components of Angiogenesis
3) CELL ADHESION MOLECULES (CAM) Mediate cell-cell adhesion processes Selectins IG Supergene family- ICAM VCAM Cadherins Integrins- vitronectin receptor 4) PROTEASES Degrade ECM to provide suitable environment for EC migration thru adjacent stroma Ex: Metalloproteinases (MMP)
Components of Angiogenesis
5) ANGIOGENESIS INHIBITORS Interferon TSP-1 Angiostatin Endostatin Vasostatin CLINICAL SIGNIFICANCE: Tumor angiogenesis switch is triggered as a result of shift in the balance of stimulators to inhibitors
ANGIOGENESIS
Therapeutic Implications
Metalloproteinase inhibitors (MMPI) block the degradation of basement membrane Inhibitors of endothelial functionthalidomide, TNP 470,endostatin Anti-angiogenic factors tyrosine kinase inhibitors of VEGF bFGF PDGF Interferon angiogenic inhibitor COX-2 inhibitor thromboxane A2 as critical intermediary of angiogenesis
PROCESS OF METASTASIS
Triad of Invasion
Adhesion with the basement membrane. Local proteolysis Mobility and ability to translocate through rents in bodys structural barriers.
ADHESION
De-regulated function of CAM (E-cadherin) Changes in catenin expression leads to loss of cadherin function Integrin over expression in naturally occurring cancers Downregulation of integrin in more advanced stages of cancer Upregulation of ICAM-1 which enhances extravasation Adhesion molecules on EC: E-selectin,VCAM
ICAM
LOCAL PROTEOLYSIS
Degradation of basement membrane to traverse barriers Carried out by: 1. Serine proteases -uPA elastase plasmin cathepsin G 2. Cysteine proteases- cathepsin B L 3. Aspartate proteases cathepsin D 4. Matrix metalloproteinasesgelatinases interstitial collagenases stromelysins matrilysins
MOTILITY
Tumor cells can move randomly or directionally toward attractants Modulators of motility
GF, hyaluronases, components of ECM, tumor-secreted factors, host-derived factors
Tumor p53 suppressor protein is reffered to as guardian of the genome bec. it:
A. Enhances the survival of tissues B. Allows apoptosis to occur on seriously damaged cells C. Plays a key role in G2 checkpoint control D. Arrests the cell cycle at Go phase
Tumor p53 suppressor protein is reffered to as guardian of the genome bec. it:
A. Enhances the survival of tissues B. Allows apoptosis to occur on seriously damaged cells C. Plays a key role in G2 checkpoint control D. Arrests the cell cycle at Go phase
Any Questions ?