Melanoma Pathology,

Treatment and Genetics

 Key Issues
• Incidence

• What is melanoma?
– Pathology

• Pathology/Aetiology of Skin cancer.

– Causes
– Who gets it
– Location of melanomas (what part of body)
– Role of UV radiation

• Treatment of Melanoma

• Melanoma Associated genes

– Tumour suppressor genes
– Oncogenes

• Antigenicity of Melanomas.

• A genetic model of melanoma.

 Incidence of Melanoma

• Skin cancer is the second most common cancer in the

UK.
– About 50,000 cases per year in the UK (includes SCC
& BCC)
– 5-6000 are melanoma

• 1 in 80 people in the US/Australia get CMM

• 1 in 200-250 in the UK get CMM

• CMM is responsible for 90% of skin cancer associated

deaths
 Cutaneous Malignant
Melanoma
• CMMs are derived from the melanocytes located in
the basal layer of the epidermis.

• Very aggressive tumour and metastasise to a variety

of organs
– Bone
– Brain
– Juxtaposed lymph nodes

• Two modes of growth

– Radial growth phase – over surface of skin
– Vertical growth phase – into lower layers prior to
metastasis
Skin Anatomy
 Melanoma Types

Lentigo MM Superficial Spreading MM

Acral MM Nodular MM
Melanoma Pathology
 Cutaneous Malignant
Melanoma
• If tumour resected before a vertical invasion of less
than 3mm - associated with a cure.

• Greater than 3mm – metastatic disease likely.

• 80% of CMM patients get metastatic disease.

• Metastatic lesions associated with about 6 months

survival following diagnosis.
Local Spreading of Melanoma

Radial Growth Phase

Vertical growth phase

Malignant
Point of origin: Acquired/dysplastic neavus (mole)
 What Causes Melanoma
• Strong evidence that melanoma occurs on sun
damaged skin.

• High risk associated with short bursts of intense UV

exposure.
– Intermittent exposure during childhood - important factor.

• Who gets it
– Those with high number of naevae (moles).
– Type 1 skin – blue eyes, blond, burn easily, those
with a family history of melanoma.

• Age group: 40 –60 year olds, but can occur at any age
(rarely in children)

• More women than men get melanoma.

Pathology/Aetiology of
Skin Cancer
 
SCC BCC CMM

Sun Sensitive skin • • •

 
 

Total Sun Exposure • •

 

 
 

• •
 
Childhood Sun
Exposure
 
• •
 
Intermittent Over
Exposure
• •
 
P53 Mutation
 
 Location
• Affect most parts of the body.
– Women – legs.
– Men – trunk especially the back.

• Role of Sunlight (UV irradiation)

– UVA: 320 – 400nm
– UVB: 280 – 320nm
– UVC: 100 – 280nm

• All UVC and some UVB is blocked by atmospheric ozone.

• UVB causes the erythemal response – reddening associated with sun

burn.

• UVA: Skin ageing

 Treatment of Melanoma
• If melanoma is detected early:
– Surgical resection of primary tumour followed by
careful monitoring.

• Advanced Disease
– Resection of Primary tumour
– Lymph node excision and surgical removal of
metastases where possible
– High dose interferon to treat disseminated
disease (boost immune system)
– Melphalan + Interferon

• Prognosis is poor with advanced disease

 Experimental Models
• Xiphophorus fish: Irradiation with single dose UVB
induces melanoma in fish (Setlow).
– Spectral regions contributing to melanoma: a personal view.
Setlow RB, J Investig Dermatol Symp Proc. 1999 Sep;4(1):46-9

• XPA transgenic mice: Knockout mice (excision repair

gene XPA) are susceptible to melanoma induction by
UVB irradiation
– Photocarcinogenesis: UVA vs UVB.
de Gruijl FR.Methods Enzymol. 2000;319:359-66.

• Aim is to produce an Action Spectrum in humans

Determination of what UV wavelength is responsible
for CMM induction.
 Molecular Genetics of
Melanoma
• A number of chromosomal aberrations are known
to accumulate during CMM development.

• Principally non-random Losses of Heterozygosity

(LOH) (remember Knudson’s two hit hypothesis of
retinoblastoma)
LOH in Melanoma

Chromosome Frequency of LOH

1p36 25%
6q22-24 30-35%
9p21 (p16) 50-60%
10q22-25 30%
11q23-ter 25%
 
Tumour Suppressor Genes

• No evidence of P53 involvement in melanoma development.

• CDKN2A/INK4A/P16: Cyclin dependent kinase 4 inhibitor.

• Causes a G1 arrest by inhibiting the phosphorylation of RB.

Inhibits the action of cyclin dependent kinases 4 and 6.

• Gene Structure:
– 3 exons
• Exon 1: 150bp (50aa)
• Exon 2: 307bp (101aa)
• Exon 3:12bp (3 aa)
 Cell Cycle Control
(abbrev.)

CyclinD1

G0 CyclinD2 Cyclin E Cyclin A Cyclin B

CyclinD3
pRB
G1 S G2 M

CDK6 CDK2 CDK2 CDK1

CDK4
p15, p16, p18, p19
The INK4 locus of chromosome 9p21

Exon 1 Exon 2 Exon 1β Exon 1α Exon 2 Exon 3

p15 Exon 1 Exon 2

p16 Exon 1α Exon 2 Exon 3

p14 ARF Exon 1β Exon 2 Exon 3

 Inactivation of P16

• Deletion: About 50-60% of melanomas display LOH at

chromosome 9p21 (location of P16).

• P16 loss is thought to be a critical event in

melanogenesis.

• Methylation: CpG islands in the promoter region of

P16. Causes functional silencing of the gene.
– Occurs frequently in head and neck (SCC), breast,
prostate, bladder cancers.
– Probably a significant event in melanoma development

• Point mutation: Observed in many cancers including

melanomas
 Point mutations in P16

• Inactivation of P16 by point mutations not a result

of prior exposure to UV irradiation.

• UV exposure = pyrimidine dimers in DNA.

• Mutation by UV = CC→TT tandem base substitution.

• Never observed in any known melanoma tumour

suppressor gene.
– Despite the obvious role of UV in melanogenesis.

• Mutation of P16 in the germline of melanoma

kindreds establishes P16 as a familial TSG.
Pyrimidine Dimer
Other Melanoma Tumour Suppressor
Genes

• PTEN: Phosphatase and Tensin homologue.

– also called
• MMAC1: Mutated in Multiple Advanced Cancers.

• Encodes a tyrosine phosphatase with extensive homology

to the cytoskeletal proteins auxillin and tensin.
– Suppresses Akt signalling

• PTEN/MMAC1 maps to chromosome 10q23.3

– 9 exons
– ORF: 1212nt
– 403 amino acids
– 47kDa

• Functions as a negative regulator of signal transduction

 Any Other Melanoma Tumour
Suppressors?

• Complex LOH pattern during melanoma development.

– Likely that other (as yet undiscovered) TSGs are
inactivated during melanogenesis.

• Chromosome 9:
– Pollock et al, 2000. Evidence for three tumor
suppressor loci on chromosome 9p involved in
melanoma development. Cancer Research. 61: 1154-
1161.
– Parris et al., 1999. Functional Evidence novel
tumour suppressor genes on chromosome 9. Cancer
Research. 59: 516-520.

• In many melanomas P16 is left intact and deletion

occur next to P16 on chromosome 9p21
 Any Other Melanoma Tumour
Suppressors?
• KiSS1 Tumour suppressor Gene
– Chromosome 1p32
– Regulates an antimetastasis gene on
chromosome 6.
– Associated with LOH on chromosome 6q16

• LOH also demonstrated on

– chromosomes 11
– chromosome 10 (not PTEN)

• As yet most of the tumour suppressor genes have

not been fully identified.
 Oncogene Activation and Melanoma

• N-RAS mutational activation at codon 61

– CAA→AAA

• Destroys natural ATPase activity of Ras protein.

• Observed in
– Congenital naevi
– Dysplastic naevi
 Oncogenes

• B-Raf oncogenes: Ras-Raf MAP kinase signal

transduction pathway.

• A serine threonine kinase leading to

upregulation of BRN-2 transcription factor

• Mutations in 60% of primary melanomas

• Elevated kinase activity

– Capable of transforming NIH3T3 cells in
culture.
Other Potential Melanoma Associated
Genes
• Integrins

• A diverse family of receptors that mediate adhesion

between the cell membrane and the extracellular matrix.

• Composed of α and β subunits.

• Combinations of 14 α and 8 β subunits comprise a

family of over 20 integrin types.

• Certain classes of integrin expression has been shown to

be upregulated during transition from radial to vertical
growth phase.
– Integrin α vβ
 Antigenicity of Melanomas
• Melanomas are thought to have an antigenic
component
– Photo-immunological aspects of melanoma.
J.Investigative Dermatology. 24: 153-165
(1995) Kripke ML et al.

• May render them sensitive to immunological

methods of treatment.

• High molecular weight melanoma associated

antigen: associated with many melanomas

• Possible exploitation for immunotherapy.

 A Genetic Model

Chromosomes 6
and 11
N-Ras Chromosome 9 Integrins? Chromosome 1(KiSS 1)
P16/other TSGs and 10 (PTEN/MMAC1)

Normal melanocyte Dysplastic or Radial Vertical Metastatic

acquired naevus growth phase growth phase Melanoma

B-Raf
Chromosome 7q34
 Summary
• Incidence

• What is melanoma?
– Pathology

• Pathology/Aetiology of Skin cancer.

– Causes
– Who gets it
– Location of melanomas (what part of body)
– Role of UV radiation

• Treatment of Melanoma

• Melanoma Associated genes

– Tumour suppressor genes
– Oncogenes

• Antigenicity of Melanomas.

• A genetic model of melanoma.