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Investigating Cancer

KRAS Activity

What is cancer?
All cancers derive from single
cells that have acquired the
characteristics of continually
dividing in an unrestrained
manner and invading
surrounding tissues.
Cancer cells behave in this
abnormal manner because of
changes in the DNA sequence
of key genes, which are known
as cancer genes. Therefore all
cancers are genetic diseases.

Human melanoma cell undergoing cell


division
Credit: Paul Smith & Rachel Errington, Wellcome Images

Cancer information
One in three people in the Western world develop
cancer and one in five die of the disease
There are approximately 200 types of cancer, each
with different causes, symptoms and treatments
In 2007, 297,991 people were newly diagnosed with
cancer in the UK
An individual's risk of developing cancer depends on
many factors, including age, lifestyle and genetic
make-up
Cancer Research UK
http://info.cancerresearchuk.org/cancerstats/incidence/?a=5441

The 20 most common causes of


death from cancer, UK, 2008

Cancer Research UK. Accessed July 2010


http://info.cancerresearchuk.org/cancerstats/mortality/cancerdeaths /

Cancer cells have altered


genomes
Karyotype illustrating structural abnormalities in cancer

Credit : Mira Grigorova and Paul Edwards, Department of Pathology, University of Cambridge, unpublished
Source: www.path.cam.ac.uk/~pawefish/BreastCellLineDescriptions/HCC38.html

What is a mutation?
Germline mutation
A change in the DNA sequence that
can be inherited from either parent

Somatic mutation
A change in the DNA sequence in cells
other than sperm or egg
The mutation is present in the cancer
cell and its offspring, but not in the
patients healthy cells

Mutations &
cancer genes
Cancer genes are causally implicated in
oncogenesis
Mutations in cancer genes can occur somatically
or can be inherited.
Mutations in some cancer genes can be inherited
from parents, in which case they are present in
every cell of the body. Such people are at a
higher risk of developing cancer.
Somatic mutations can occur in any of the cells of
the body except the germ cells (sperm and egg)
and therefore are not passed on to children.

Importance of somatic
DNA changes in human
Both
cancerInherited
Somatic

Only 5 10% of cancer cases have a clear hereditary component,


e.g. BRCA1 and BRCA2 in breast cancer
Even in those cases where susceptibility is clearly inherited, somatic
changes are required for cancer to develop

Cancer genes
There are two types of cancer genes:
Tumour suppressor genes
Oncogenes

To date, we know of approximately 400 somatic


cancer genes * but there are almost certainly
more to be found
COSMIC is a catalogue of somatic mutations
found in cancer genes in human tumours and is
available at:
http://www.sanger.ac.uk/genetics/CGP/cosmic/
*(COSMIC v47release. July 2010)

Tumour suppressor gene


These genes normally function to PREVENT
cell growth/division

TS

Cancer

Oncogene
Genes which normally function to PROMOTE cell
growth/division in a controlled manner
Ras

Cancer

Examples of mutations
Sequence
1

ACTCGTTAGGCA

Sequence 2

Type
Substitution

ACTCGTTAGGCA

ACTCCTTAGGC
A
ACTCGGCA

ACTCGTTAGGCA

ACTCGTTATCAGGCA

Insertion

ACTCGTTAGGCA

ACTTTGCAGGCA

Inversion

ACTCGTTAGGCA

ACTCGTTAGTTAGGCA

Deletion

Duplication

Cancer progression
Mutations in multiple cancer genes are required for the
development and progression of a single cancer

Benign
Tumour
In situ
cancer
Invasive
cancer
Metastatic
cancer

External causes of cancer:


ultraviolet radiation

www. flickr.com: lastexit

External causes of cancer:


tobacco smoke

Lifestyle factor: diet

Biological factor: virus


HPV is a cause of
cervical cancer
Proteins from the virus
activate and deactivate
cancer genes
The role of HPV in
cervical cancer has led
to the development of
vaccines
HPV in cervical epithelium
Credit: MRC NIMR, Wellcome Images

Activity
The KRAS gene codes for a signalling molecule
Mutations in KRAS are present in many
cancers, including pancreatic cancer
You have to look for the mutations by
comparing healthy DNA sequence with tumour
DNA sequence
Not all of you will find a mutation

Your Worksheets

If you find a
mutation
EX
AM
PL
EO
NL
Y

How to use the codon


wheel
Start from the
centre and move
outwards

Mark up your sequence

Heterozygous mutations
Normal DNA
sequence

DNA change
in cancer
T A
BRAF gene mutation Nature 417, 906-7 (June 2002)

A double peak
indicates a mutation
on one chromosome
and not the other i.e.
a heterozygous
mutation

Results
Amino
Acid
Number
Acid
Number

12
12
13
13
30
30
61
61
146
146
173
173

Healthy DNATumour DNA Healthy AminoTumour


Amino
Sequence DNA
Sequence
Acid Acid
Acid Acid
DNA
Amino
Amino
Sequence Sequence
G (glycine
)
V (valine
)

GGT
GGT
GGC
GGC
GAC
GAC
CAA
CAA
GCA
GCA
GAT
GAT

GTT
GTT
GAC
GAC
GAT
GAT
CGA
CGA
CCA
CCA
GAC
GAC

G (glycine)
V (valine)
G (glycine)
D(aspartic
acid)
G (glycine)
D (aspartic
D(aspartic
acid) acid)
D(aspartic acid
)
D (aspartic
D (Aspartic
Q (glutamine
)
R (arginine
)
acid)
acid)
Q
R (arginine)
A (alanine)
P (proline)
(glutamine)
A (alanine)
P (proline)
D(aspartic acid
) D (aspartic
acid)
D (Aspartic
D (Aspartic
acid)
acid)

Significant mutations
Amino Acid Healthy
Number
DNA
Sequence

Tumour
DNA
Sequence

Healthy
Tumour
Amino Acid Amino Acid

12

GGT

GTT

G (glycine)

V (valine)

13

GGC

GAC

G (glycine)

D (asparatic

61

CAA

CGA

146

GCA

CCA

A (alanine)

acid)

R (arginine)

(glutamine)

P (proline)

How common?
AA 12
13,894

AA 13
2,111

AA 61
212

AA 146
33

Source: COSMIC July


2010

RB1: tumour suppressor


gene

Source: COSMIC July


2010

How does this affect the


KRAS protein?

Amino acid 12

Amino acid 13

Amino acid 61

Amino acid 146

Amino acid 146

Whats the impact?


KRAS helps to transmit external growth signals to the
cell nucleus, driving normal cell growth. It is:
Activated when it binds GTP
Inactivated or switched off when GTP is
hydrolysed to GDP