RADT 112

-1-

UNIVERSITY OF OTAGO EXAMINATIONS 2015

Examination for the Degree of

Bachelor of Radiation Therapy

Paper: RADT 112

CANCER CELL BIOLOGY

Date & Time: Friday, 5th June 2015

9.00am to 11.00am

Time Allowed: TWO (2) Hours

Room Number: RT22

Marks: TOTAL: 100 Marks

This examination accounts for 40% of the available marks for
the paper.

INSTRUCTIONS: Answer ALL questions.

Answer each question on a new page.
Ensure you read each question carefully.
All diagrams should be clear and well labelled.
Write your student ID Number on each answer book used.
Use separate answer books for Part A and Part B.

This examination paper comprises three (3) pages

PART A: ANSWER ALL QUESTIONS

1. Explain the following quote: “Cancer is a multistage process”.

 RADT 112
-2-

-Cancer involves an accumulation of changes in gene expression eg. Mutations that cause
dysregulation of functional characteristics

-these are tightly regulated as genes require addition of functional proteins to be activated
and thus processes should only occur if they are needed.

- normal cells will only undergo transcription if it is sure the right gene is there and is needed
to be copied, but cancer cells have a mutation that may cause signalling pathways to be
initiated unnecessarily.

- dysplasia then occurs which is the increase in number of abnormal cells that can then lead to
the formation of a tumour mass with malignant growth.

- free radical formation can also occur where a highly reactive molecule with an unpaired
outer electron can cause a cascade of reactions as it breaks bonds and damages DNA further.

- the DNA repair mechanisms in cancer cells are less efficient than normal cells and thus
accumulates many mutations without repairing them

- cancer requires many mutations, ie. PO genes may be upregulated to increase the growth
ability/ signals. Then tumour suppressor genes may be suppressed (dysregulated) to prevent
their action to prevent tumour formation

Total: 5 Marks

2. (a) Explain what is meant by “the epigenetic profile of a cell”? (3 marks)

Epigenetic profile is what genes are expressed or suppressed, the epigenome is DNA wrapped
around histones and covered in chemical tags. The epigenome is influenced by endogenous or
exogenous signals to the surrounding environment. It can wind or unwind in order to make
genes accessible or non accessible and thus determines if they are expressed or not.

(b) Explain how the epigenetic profile of cancer cells differs from that of normal cells. (3
marks)

Normal cells regulate transcriptions so that it only occurs if needed and to the correct genes. Cancer cells
accumulate changes in gene expression ie. Genetic profile, which alters functional characteristics. Mutations
within cancer cells cause dysregulation of proteins that may mean upregulation or over expression of factors
to increase proliferation. The epigenetic profile expresses and silences different genes in order to sustain
proliferation and resist cell death to become malignant and anaplastic.

Total: 6 Marks

3. (a) Draw and label the cell cycle and explain briefly the processes that occur in
each of these phases. (8 marks)
G0- most resistant to RT as non proliferating,
G1- cell is active
 RADT 112
-3-

S- synthesis phase where DNA replication occurs (helicase unwinds DNA and DNA polymerase reads
and transcribes a new strand.
G2- cell growth continues
M- mitosis, prophase (centrosome separates and chromosomes condense and NEB, prometaphase
(spindle formation), metaphase (chromosomes align), anaphase (chromosomes separate), telophase
(cytokinesis- nuclear envelope reformed and 2 daughter cells result)

(b) Name and explain the importance of the cell cycle check points.
- Cell cycle checkpoints are at G1/S and G2/M phases in the cells cycle, they are important to prevent
mutations or damage to DNA continuing to mitosis and getting passed on to the daughter cells.
- EGF plays an important role in initiating the signal cascade to drive the cell cycle to proliferation. The
checkpoints ensure that there is no dysregulated proteins that will lead to unnatural cells and possible mitotic
catastrophe.
- the checkpoints allow the cycle to stop if damage is detected and repair can then occur or induced
apoptosis, EGFR can be over expressed and thus cause increased proliferation, the cycle checkpoints are
important to detect abnormality and stop the cycle if needed.
(4 marks)

Total: 12 Marks

3. Describe three different factors that contribute to differences in drug reaction between
individuals.
Co administration= limited capacity of metabolic enzymes may mean that taking another drug at the
same time will reduce the metabolism and effect resulting from the drug. This can also increase
toxicity.
Disease status= impaired function or co morbidities ie. Hepatic dysfunction will impact the
metabolism and effect of the drug as it absorbed and into the systemic circulation
Pharmacogenetics= some genetic differences ir. Mutations may increase or decrease the metabolic
capacity. And thus some people may have an enhanced metabolism to increase their response or the
timing of their response. This can include the over expression of enzymes or their receptor.

Total: 6 Marks

4. Explain what is meant by “hepatic first pass” and how can this affect the effect of the drug on
tissues.

this is relevant when a drug is being administered per oral (PO) as the pharmacokinetics (what the
body does to the drug) involves a system of different processes to get the drug to its target. This is
influenced by absorption into blood vessels, distribution round the body, metabolism in the liver and
excretion through sweat, urine, breast milk etc.
The first pass effect is the impact of how bioavailability is decreased as the drug must diffuse
endothelium and be absorbed into the blood stream. The drug enters the hepatic portal system
where it goes to the liver to be metabolised. This then means some of the drug is transferred into
active metabolites and there is less drug available to interact with the target.
This means that drugs administered orally are usually of a higher dose as they have to account for the
loss in bioavailability (unchanged drug that will interact with the target) compared to intravenously
 RADT 112
-4-

administered drugs that are injected straight into the blood stream and the full does reaches the
systemic circulation to be delivered to the target.

Total: 6 Marks

6. (a) Compare and contrast cytotoxic and targeted chemotherapies (8 marks)

Cytotoxic chemo- kills all fast proliferating cells (including normal cells), interferes with DNA function
and cell division,
Targeted chemo- kills only cells with a specific molecular target, inhibits proliferation pathways,

Both therapies aim to inhibit the proliferation of cancer cells and lead to increased apoptosis and
decreased survival, and invasion/metastases

(b) Name an example of each and name one type of cancer it is used for (2 marks)
Cytotoxic- alkylating agent- cisplatin- reacts with bases in DNA and causes crosslinking and prevents
gene expression/ replication and transcription- bladder cancer
targeted therapy- monoclonal antibodies- trastuzumab- targets the external domain of the HER2
receptor and prevents dimerization and initiation of the signalling cascade.
Total: 10 Marks

7. Describe the mode of actions of a named mitotic inhibitor and a named topoisomerase
inhibitor as anti-cancer drugs.

paclitaxel- binds to tubulin and stabilises actin filaments to prevent spindle formation and
prevents mitosis occurring properly. This occurs in M phase and is for breast and ovarian
cancers
etoposide- prevents the action of topoisomerase which relaxes and recoils DNA to allow DNA
replication to occur. Occurs in S phase and is for testicular cancer, block the ability to reanneal
after cuts are made to the DNA strand. Kills proliferating and cells with DNA damage or
actively metabolising cells.
Total: 6 Marks

8. Describe the mode of action of vemurafenib and explain in which instance it would be used as
an anti-cancer agent.
Total: 6 Marks
Targets the BRAFv600e mutation, targets intracellular to prevent the mutation causing signal
cascade. This binds to and blocks the protein kinase to prevent out of control proliferation, in turn up
regulating apoptosis and decreasing survival and invasion. This is in melanoma and has shown to have
an initial good response but the tumours re-establish over time and return to be resistant to the drug.

Part A subtotal: 57 Marks

PART B: ANSWER ALL QUESTIONS (USE A SEPARATE ANSWER BOOK FROM PART A)

9. Explain how the ionisation cascade differs from the free radical cascade in the context of
radiation therapy.
 RADT 112
-5-

RT involves photons causing the release of high energy electron from the outer shell of an
atom of molecules in the human body. This electron can then collide with other outer shell
electrons and cause them to further react and a cascade of ionisation occurs. The ionisation
cascade is the physical phase.
The free radical cascade is the chemical phase that follows the physical phase. This involves the
electrons causing damage to organic macromolecules (specifically DNA damage and DSB). The
electrons lose energy as they collide and eventually combine with an atom/molecule and
form a free radical. These are then highly reactive as they have an unpaired electron in the
outer shell that will cause it to break bonds to cause DNA damage such as single or double
strand breaks. This is then a cascade as affected atoms then go on to react further because
they now have an unpaired electron and aim to get to a more stable state.
Photons can directly ionise DNA and cause SSB or DSB but it is primarily from the formation of free radicals
that react with surrounding atoms in human tissue. As the body is 70% water, the oxygen is likely to react
from radiation therapy and form a ROS (free radical).
Total: 8 Marks

10. Describe the following processes of cell death.

a. Apoptosis- suicide, cell forms blebs, nucleus condenses, whole cell eventually
disintegrates and is disposed of by macrophages. Initiated by receptor on membrane that
detects damage and checkpoints stops cell cycle to try repair and if not possible-
apoptosis.
b. Mitotic catastrophe- most common form of cell death in radiation therapy, cells with
damage enter mitosis and are unable to complete it. They may have two centromeres
and result in large multi nucleated cells that do not function properly. They cannot divide
and then end in cell death.
c. Necrosis – injury, cellular swelling and membrane breakdown and organelle breakdown
lead to death as cell cannot function properly, this affects large numbers of cells and is
shown by inflammation
d. Senescence- as DNA replication occurs, the telomeres (non coding regions at the end of
DNA strands) are slowly shortened and eventually gone. This equals death via
‘senescence’ as the telomeres are required by the cell to ensure no important coding
DNA is lost.
Total: 9 Marks

11. Within the context of the linear quadratic model, explain what αD and βD 2 represent.
alpha is the linear component of the model, where single hits cause double strand breaks as
alpha particle are less penetrating but more damaging. This is the component where most cell
death is due to alpha and has a steeper gradient as it occurs more at low doses.

beta is the quadratic component, where a larger proportion of the dose contributes to cell
death via multiple single strand breaks from beta particle. This is more common at higher
doses where single strand breaks are more likely to occur closer together and cause a double
strand break.
The linear quadratic model displays total cell death from both of the particles, the alpha beta
ratio is the point at which cell death is equal from both alpha and beta particle.
 RADT 112
-6-

Total: 4 Marks

12. Describe how a tumour’s α/β ratio might influence the use of a non-standard
fractionation regime.

- A higher ratio is in early responding tissues

- We know lower doses for an increased number of fractions is appropriate as the cells are
relatively more sensitive at lower doses (hyperfractionation).
- High ratio, early responding tissues are also usually fast proliferating and thus the increased
number of fractions leaves the cells less time to repair their damage.
- High ratio early response tissues also have a large amount of stem cells that are more
radioresistant so multiple treatments required to treat them
- Low ratio tissues are late responding and are slow at proliferating,
- they are also relatively more sensitive at higher doses so a hypofractionation regime can be used
with increased dosage in a smaller number of fractions.
- This will overwhelm the cells repair mechanisms as it has a smaller amount of stem cells and
reduced mitotic turnover
Total: 8 Marks

13. Describe the mode of action of a named anti-oestrogen, and the type of cancer it can
be used to treat.

tamoxifen- ER positive breast cancer- targets the ER to prevent dimerization, it binds to the
receptor but does not activate the gene and thus no downstream signal cascade.

This interferes with the hypothalamus, pituitary, ovaries axis that leads to the production of
oestrogen. [Diagram]
The hypothalamus releases GnRH and receptors on the pituitary signal the release of FSH and
LH that go to receptors on the ovaries that then produces oestrogen. Tamoxifen binds to ER in
the intracellular network of the ovaries so that oestrogen can’t bind and there is no signal
cascade in the cell to initiate proliferation. This is a Selective oestrogen receptor modulators
as it acts as an anti estrogen in some tissues and estrogen like in others. This is for
menopausal women as they produce a small amount of estrogen that will then be blocked in
the cancer cells. This is less effective for pre-menopausal women (unless taking goserelin) as
they produce high levels of estrogen in the ovaries.
Total: 8 Marks

14. (a) Name the cancer diagnosis for which abiraterone is funded.
castration resistant prostate cancer
(b) Describe abiraterone’s mode of action, and explain why it is an effective anti-cancer
agent for this diagnosis.
abiraterone is a testosterone biosynthesis inhibitor that targets CYP17 and blocks its
activity. This is a form of androgen deprivation therapy as it prevents all synthesis of
testosterone on the testes and the adrenal glands, thus no testosterone to enhance
proliferation of hormone receptor positive prostate cancer. This is only in advanced
cases where patients have not had a successful response from previous hormone
 RADT 112
-7-

treatments, as it can be toxic to organs. This is different to bicalutamide as it prevents

all synthesis of testosterone and not just prevent the binding to receptors.
Total: 6 Marks

Part B subtotal: 43 marks

TOTAL FOR EXAMINATION: 100 MARKS