Neoplasia

By
Prof. J.T. Anim
Department of Pathology
Lecture IV
Topics
 The neoplastic cell
 Causes of neoplasia
 Pathogenesis/mechanisms of
neoplasia - oncogenesis
 Carcinogens and carcinogenesis
Biology of Neoplastic Cell
 Morphological Changes
 High nucleo-cytoplasmic ratio
 Irregular nuclear shape
 Prominent or multiple nucleoli
 Numerous protrusions on cell surface
 Disorganised cytoskeleton
Biology of Neoplastic cell
 Biochemical alterations
 Composition of cell surface
 Loss of glycoprotein molecules
 Deletion of saccharide residues from glycolipids
 Loss of fibronectin
 Cell surface properties
 Enhanced mobility of receptors resulting from loss of cytoskeletal
attachment to integral glycoproteins
 Expression of foetal, viral, or neo-antigens
 Shedding of antigens (and other glycoproteins)
 Increase in net negative charge on plasma membrane
 Enzyme production and release
 Increased protease activity (including collagenase and pl. activator)
 Increased release of alkaline phosphatase and glycosyl transferase
 Transport – increased uptake of sugars and amino acids
 Cyclic nucleotides – increased cyclic GMP
Biology of Neoplastic Cell
 Selection
 Selective pressures confer advantages
on some tumour cells → progressively
more homogeneous population.
 The host immune system
 Limits of space and nutrition
 Lethal mutations
 Post-mitotic arrest
Biology of Neoplastic Cell
 Growth and division - modified by:
 Intracellular factors
 Polyamines, eg. spermidine, released in dividing cells
 Cyclic AMP inhibits cell growth
 Protein kinases control progression of cell cycle by regulating phosphorylation
of histones
 Extracellular factors
 Nutrients
 Calcium ions
 Growth stimulating hormones and polypeptides - insulin stimulation or specific
stimulation by:
 Oestrogens – some breast and endometrial carcinomas
 Epidermal growth factor
 Platelet derived growth factor
 Sarcoma growth factors, etc
 Cancer-associated galactosyl transferase acceptor (CAGA) inhibits growth of
neoplastic cells
 Others – modify environment of cancer cells
 Proteolytic enzymes reduce physical resistance by surrounding tissues
 Angiogenesis factors stimulate new blood vessel formation and nutrition
Biology of Neoplastic Cell
 Behavioural characteristics in vitro
 Immortality – can proliferate ad infinitum under ideal
culture conditions
 Loss of contact inhibition – proliferation persists
 Increased motility – migrate more rapidly (may be due to
altered cytoskeletal function)
 Decreased cell-to-cell adhesion – disturbances in
arrangement of macromolecules on tumour cells
 Loss of substrate dependency – show a greatly
diminished dependence on substrate attachment through
binding proteins, eg. fibronectin, laminin
Aetiology of Tumours
 The sequence of intra and
extracellular mechanisms are
unknown
 Genetic mutation and/or altered gene
expression are critical factors in
oncogenesis
 Genetic factors interact with
environmental factors
The Transformed Cell
 Autonomous generation of mitogenic signals
 Insensitivity to exogenous antigrowth signals
 Resistance to apoptosis
 Limitless replicative potential
(immortalisation)
 Blocked differentiation
 Ability to sustain angiogenesis
 Capacity to invade surrounding tissues
 Potential to metastasise
Pathogenesis of Neoplasia
 Non-lethal genetic damage
 Mutation may be inherited in the germ line or
acquired somatic (chemical, radiation, viruses)
 Leads to clonal expansion
 Role of regulatory genes
 Oncogenes and tumour suppressor genes
 Genes regulating apoptosis
 Genotypic and phenotypic multistep process
 Stepwise acquisition of characteristics of a
neoplasm – tumour progression
Oncogenesis
 Oncogenes
 Cellular oncogenes – tumour promoting genes derived from
normal growth promoting genes in normal cells (proto-
oncogenes)
 Viral oncogenes (v-oncs) – a family of transforming genes, each
of which is characteristic of a rapidly transforming oncornavirus
 Tumour suppressor genes
 Growth inhibiting genes – encode negative transcriptional
regulators of cell cycle, signal transducing molecules, cell surface
receptors
 Mutator genes (caretaker genes)
 DNA mismatch repair genes – exercise surveillance over integrity
of the genetic information by participating in cellular responses to
DNA damage. Loss of their function → DNA susceptible to
progressive accumulation of mutations
Proto-oncogenes
 Growth factors – sis, int-2, IGF-1
 Growth factor receptors with protein kinase – erb-2, fms, met, trk, ret
 Abnormally functioning growth factor receptors – neu
 Factors that act in the transduction of signals arising from ligand-
receptor interactions
 G proteins – ras
 Guanosine 5-triphosphatase activators (GTPase) – gap, brev
 Membrane-associated cytoplasmic kinases – src, yes, fgr
 Non-membrane associated cytoplasmic kinases – raf, mos, pim-
1, fps
 DNA-binding proteins concerned in transcription
 Heterodimeric transcription factors – fos-jun, myc-max
 Transcription factors – myb, rel, ets
 Cell cycle proteins - cyclins
 • Growth factors
• Transmembrane growth factor
receptors (tyrosine kinase)
• Membrane associated kinases
• ras GTPase family
• Cytoplasmic kinases
• Nuclear transcriptional factors
Cellular compartments in which oncogene or protooncogene products reside
Gene products of protooncogenes and steps involved in cell division
Activation of Cellular Oncogenes

 Mutation of proto-oncogene
 Leads to constitutively activated
abnormal protein
 Increased expression of the proto-
oncogene
 Overproduction of a normal gene product
 Insensitivity of normal auto- inhibitory
and regulatory constraints
Mechanism of activation - mutation
Mechanism of activation - translocation
Mechanism of activation - amplification
Mechanism of activation
Summary of oncogene activation
Mechanism of Oncogene Action

 Growth factors
 Cell surface receptors
 Intracellular signal transduction pathways
 DNA-binding nuclear proteins (transcription
factors)
 Cell cycle proteins (cyclins and cyclin-
dependent protein kinases)
 Inhibitors of apoptosis (bcl-2)
Viral Oncogenes
 Viral oncogenes originate from cellular
proto-oncogenes
 The current view is that v-oncs arise
as a result of transcription from a
cellular proto-oncogene, and that this
cellular genetic material has been
incorporated into the viral genome
Viral Oncogenes
 A number of v-oncs code for proteins that
possess the ability to phosphorylate tyrosine
kinase in certain cells
 Oncogene from the avian erythroblastosis
virus (erb b) codes for a protein homologous
with a portion of the cellular receptor for
epidermal growth factor.
 Another oncogene, sis from the simian
sarcoma virus, codes for a protein
homologous with PDGF
Tumour Suppressor Genes
 Encode negative transcriptional regulators
of cell cycle
 Encode signal transduction molecules
 Encode cell surface receptors
 Both alleles must be inactivated to produce
the deficit that allows tumour development
 Heterozygous state is sufficient to protect
against cancer
 Loss of heterozygosity predisposes to
tumour development
Tumour Suppressor Genes
 Certain hereditary tumours are
associated with mutation or loss of
alleles in germline cells
 Retinoblastoma (RB gene) 13q14
 Wilms’ tumour (WT-1) 11p13
 Neurofibromatosis type 1 (NF-1) 17q11.2
 Familial adenomatous polyposis coli
(FAPC) 5q15-22
Tumour suppressor gene – the retinoblastoma gene
Tumour Suppressor Genes
 The p53 tumour suppressor gene
 Mutation appears to be the commonest
abnormality related to tumour suppressor genes
in human neoplasms.
 Causes arrest in G1 phase in cells with
damaged DNA to allow time for excision and
repair
 p53 protein can induce apoptosis (eliminates
cells harbouring genomic abnormalities)
Mechanism of action of p53 gene
HPV and tumour suppressor genes: Both RB and p53 proteins can be inactivated
by protein products of oncogenic human papillomavirus types 16 and 18
Properties of
oncogenes and
tumour suppressor
genes
Oncogenesis - Summary
Summary of genomic
mechanisms of cancer
Oncogenesis – Multistep Process
 Initiation
 Change in genome of target cell. If not repaired, is
permanent and heritable
 Rapid, dose-related, affects small proportion of target cells
 Promotion
 Biochemical events in normal and initiated cells → altered
pattern of gene expression → expression of cells with new
phenotype (preneoplastic cells)
 Continued effect of promoting agent → clonal expansion
 Progression
 Metastasis
Initiation and promotion:
A multistep process
Evolution of colorectal cancer
through the adenoma-carcinoma
sequence.
Oncogenesis - Genetic
 Account for less than 5% of tumours
 Chromosomally determined syndromes
 Down’s syndrome – acute leukaemias
 Klinefelter’s syndrome – breast cancer
 Gonadal dysgenesis – gonadal tumours
 Single gene abnormalities
 Xeroderma pigmentosum (aut dom) lack of enzymes for
excision and repair of DNA
 Familial adenomatous polyposis coli and related syndromes
(aut dom) inheritance of malfunctioning APC gene
 Increased risk with no precursor syndrome
 Phaeochromocytoma – childhood form (50%) or as part of
MEN IIa (Sipple’s) and MEN IIb
Oncogenesis - Chemical
 Many chemical agents
become carcinogenic only
after conversion in the body
of the host to biologically
active forms
 The parent substance =
remote carcinogen
 Its metabolites = proximate
carcinogens
 Final molecular species
that interacts with host
DNA = ultimate carcinogen
 Most chemical carcinogens
are also mutagens
Polycyclic hydrocarbons
 1775 Percival Potts: scrotal cancer in chimney sweep’s boys
 Coal tar – polycyclic hydrocarbons
 Moderate-powerful carcinogens in this group:
 7,12-dimethylbenzanthracene
 3,4-benzpyrene
 1,2,5,6-dibenzanthracene
 3-methylcholanthrene
 Site of application affects type of neoplasm produced
 Act by binding to host cytoplasmic and nuclear
macromolecules. Are activated by oxidases to water-soluble,
more reactive epoxides
 Greater DNA binding = greater carcinogenic potential
 Hydrocarbons in cigarette smoke related to lung cancer.
Predisposition to lung cancer may be related to inducibility of
aryl hydrocarbon hydroxylase
Aromatic Amines and Azo Dyes
 Aniline dye workers in
Germany – 1895
Hydroxylation in liver –
Carcinoma of bladder aminonaphthol – detox as
glucuronide
 Later: aniline dye
Aminonaphthol released
manufacture, rubber & in bladder by β-
cable industry, glucuronidase in
manufacture of certain urothelium

paints & pigments,

textile dyeing,
laboratory work
 2-Naphthylamine and
benzidine
Nitrosamines and Nitrosamides

 Nitrosamines are remote carcinogens

 Activation into alkylating agents
 Can be formed in GIT (nitrites – nitrous acid +
amines)
 Nitrites present in pickled, salted, smoked foods
 Nitrosamides do not require enzymatic
activation
 Ethyl-nitrosourea
Direct-acting Alkylating Agents
 Can bind to DNA directly
 Mustard gas
 Propiolactone
 Cyclophosphamide
 Melphalan
 busulphan
 Their interaction with DNA – useful as
antitumour agents, but also increases risk of
other neoplasms (leukaemias, lymphomas)
Naturally Occurring & Occupational

 Aflatoxin (A flavus) – produces point mutation at

codon 249 of p53 gene – hepatocellular carcinoma
 Asbestos – mesothelioma
 Arsenic – cancer of skin, lung
 Ionizing radiation – leukaemia, lung, bone
 Nickel – lung, paranasal sinuses
 Benzene – leukaemia
 Vinyl chloride – angiosarcoma of liver
 Hardwood dust – adenocarcinoma of paranasal
sinuses
 Bischlormethylether – small cell carcinoma of lung
Oncogenesis – Physical Agents

 Ultraviolet irradiation
 BCC, SCC, melanoma
 Abnormal thymine dimers in DNA of epidermal
cells; lack of efficient excision repair of abnormal
DNA
 Ionizing radiation
 Free radical damage to target cell genome
 Foreign materials
 Certain plastics – connective tissue tumours
Oncogenesis - Hormones
 Human breast cancer
 Early menarche – late menopause
 Carcinoma of endometrium
 Oestrogen effect
 Others
 Carcinoma of prostate
 Clear cell adenocarcinoma of vagina
Oncogenesis - Viruses
 DNA oncogenic viruses - Encode proteins that bind regulatory
proteins
 Papova group
 Papillomavirus: proved to cause neoplasm – >80 types, mostly
associated with benign lesions, warts (condyloma). >20 types
associated with cancer esp. 16 and 18. Major oncoproteins E6 binds
to p53 and E7 binds to RB releasing their inhibitory effect on cell
cycle.
 Polyomaviruses mainly animal tumours (rats, mice, hamsters)
 SV40 (simian vacuolating virus – no human neoplasm)
 Herpesvirus group
 Human herpesvirus 8 - Kaposi’s sarcoma
 Epstein-Barr virus: EBV genes EBNAs and LMPs involved in
immortalization of infected B lymphocytes
 Burkitt’s lymphoma
 Nasopharyngeal carcinoma
 Hepatitis group – HBV, HCV
Oncogenesis - Viruses
 Oncogenic RNA Viruses
 Adult T cell leukaemia (Japan,
Carribbean)
 HTLV-I is tropic for CD4 T lymphocytes
(leukaemia in <5%). Oncogenic stimulation
mediated by viral transcription activation
protein tax
 HTLV-II – few cases of lymphoproliferative
disorders.