Practical 4A: Analysis and scoring of tissue sections from human tumour tissue.

Introduction:
Methods: As per manual.
Results:
Discussion:
Questions:
1)H + E stands for hematoxylin and eosin stain.
2)Pathologists must take into account the cell size, the shape of cells in the sample, the presence or
absence of dividing cells and the spread of the tumour when determining tumour grade.
3) IHC is used in the diagnosis of disease. It allows the detection of specific biomarkers and
provides insight into the vigour of disease.
4)A sample is obtained and treated with hydrogen peroxidase. This blocks the action of endogenous
peroxidases in the cells, and so filters out non-specific background binding in later steps.
An unlabelled primary antibody is added to bind with the tissue antigen. The sample is then washed
to remove unbound primary antibody. A labelled secondary antibody is now added to bind to the
primary antibody, and the sample washed again to remove unbound antibodies. The addition of 2
seperate antibodies, instead of one labelled antibody specific for the tissue antigen increases the
sensitivity of the detection technique as the secondary antibody can bind at several antigenic
locations on the primary antibody.
Diaminobenzidine (DAB) is added to the sample. The oxidation of DAB develops the sample by
production of a brown dye. Excess dye is washed out of the sample, and a haematoxylin
counterstain is added to provide the contrast which aids acuity.
5)
6) TMA stands for tissue microarray. They allow a vast number of different samples to be analysed
simultaneously following treatment.
7)
8)
9)
10)
Practical 4B: Searching the biomedical literature and associated databases.
1) At the time of writing, there are 57553 articles containing the word p53.
2) At the time of writing, there are 7159 review articles containing the word p53.

3) P53 was discovered in 1979.

4) It was originally thought that p53 was an oncogene, based on the evidence that p53
cooperated with other oncogenes in in vitro transformation assays, and p53 levels are often
higher in tumour cells than in normal cells.
5) Point mutations and allele loss are common causes of p53 inactivation.
6) P63, p73 and p53 are all encoded for by homologous genes and share biological properties.
7) Cv
8) The p53 gene is located on chromosome 17. The exact position is noted 17p13.1
9) The HER2 gene is also located on chromosome 17. The gene spans the region 17q11 to
17q12.
10) sdf
11) HER2 is also known as NEU, NGL, CD340, p185, MLN 19, Neuroblastoma derived
oncogene homologue, TKR 1, EC 2.7.10, proto-oncogene c-Erb-2, proto-oncogene Neu
andTyrosine kinase type cell surface receptor HER.
12)
13) Gf
14) The unigene accession code for p53 in Homo sapiens is Hs.654481. The protein sequence is
1
61
121
181
241
301
361

meepqsdpsv epplsqetfs dlwkllpenn vlsplpsqam ddlmlspddi eqwftedpgp

deaprmpeaa ppvapapaap tpaapapaps wplsssvpsq ktyqgsygfr lgflhsgtak
svtctyspal nkmfcqlakt cpvqlwvdst pppgtrvram aiykqsqhmt evvrrcphhe
rcsdsdglap pqhlirvegn lrveylddrn tfrhsvvvpy eppevgsdct tihynymcns
scmggmnrrp iltiitleds sgnllgrnsf evrvcacpgr drrteeenlr kkgephhelp
pgstkralpn ntssspqpkk kpldgeyftl qirgrerfem frelnealel kdaqagkepg
gsrahsshlk skkgqstsrh kklmfktegp dsd

15) The predicted pI is 6.33 and the predicted MW is 43653.18

Essay bit:
Discuss the contribution of oncogenes to cancer.
Describe the hallmarks of cancer.
Explain the process of tumour angiogenesis.
What happens during cancer metastasis?

Discuss the contribution of tumour suppressor genes to cancer.

What is the basis of treating breast cancer with anti-oestrogens and Herceptin?
What are the stages of colo-rectal cancer development?