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. Accessed July 2010 http://info. 2008 Cancer Research UK.cancerresearchuk.org/cancerstats/mortality/cancerdeaths / . UK.

cam. University of Cambridge. Department of Pathology. unpublished Source: www.Cancer cells have altered genomes Karyotype illustrating structural abnormalities in cancer Credit : Mira Grigorova and Paul Edwards.path.uk/~pawefish/BreastCellLineDescriptions/HCC38.ac.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 patient’s healthy cells .

. • 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. in which case they are present in every cell of the body.Mutations & cancer genes • Cancer genes are causally implicated in oncogenesis • Mutations in cancer genes can occur somatically or can be inherited. Such people are at a higher risk of developing cancer. • Mutations in some cancer genes can be inherited from parents.

BRCA1 and BRCA2 in breast cancer Even in those cases where susceptibility is clearly inherited.g. e. somatic changes are required for cancer to develop .Importance of somatic DNA changes in human Both cancerInherited Somatic Only 5 –10% of cancer cases have a clear hereditary component.

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.ac. July 2010) .sanger.Cancer genes • There are two types of cancer genes: – Tumour suppressor genes – Oncogenes • To date.uk/genetics/CGP/cosmic/ *(COSMIC v47release.

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 .

flickr.com: lastexit .External causes of cancer: ultraviolet radiation www.

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 .

906-7 (June 2002) A double peak indicates a mutation on one chromosome and not the other i.e.Heterozygous mutations Normal DNA sequence A DNA change in cancer T→ A BRAF gene mutation Nature 417. 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 Q 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 .

driving normal cell growth.What’s the impact? • KRAS helps to transmit external growth signals to the cell nucleus. It is: – Activated when it binds GTP – Inactivated or “switched off” when GTP is hydrolysed to GDP .