DNA Damage

H. Singh Ph.D.
NTRS 467

DNA damage
DNA damage is a major topic of research within cancer
biology. Damage not only causes cancer, it is used as a
means to cure certain cancers through radiotherapy or
chemotherapy and is also responsible for the side effects of
these treatments.
http://people.bath.ac.uk/pr1cemb/DNAdamage.htm

DNA damage
Over 74,000 damage incidences occur in DNA per cell per
day, mostly by oxidation, hydrolysis, alkylation, radiation
or toxic chemicals that can either directly damage one of
the 3 billion bases contained in DNA or create breaks in
the phosphodiester backbone that the bases sit on.
The result can be mutations in genes which are transferred
the gene product (protein). If these mutations are in genes
that normally control cell proliferation or suppress tumour
growth, the cells may start to grow uncontrollably. Cells
have therefore developed mechanisms to repair DNA
damage but when they stop working efficiently, the
number of mutations in our genome increases and cancer
can develop.

Agents that Damage DNA

Certain wavelengths of radiation
ionizing radiation such as gamma rays and x-rays
ultraviolet rays, especially the UV-C rays (~260 nm)
that are absorbed strongly by DNA but also the
longer-wavelength UV-B that penetrates the ozone
shield .

Highly-reactive oxygen radicals produced

during normal cellular respiration as well as by
other biochemical pathways.

Who is responsible?
Major cause
Reactive Oxygen Species (ROS)
Reactive Nitrogen Species (RNS)

Free radicals
Superoxide

O2-

Hydroxyl

HO

Hydroperoxyl

HO2

Hydrogen peroxide

H2O2

Lipid peroxide

LO2H

Reactive nitrogen species

Thiyl

http://www.benbest.com/lifeext/aging.html#radical

Types of DNA Damage

All four of the bases in DNA (A, T, C, G) can be
covalently modified at various positions.
One of the most frequent is the loss of an amino
group ("deamination") resulting, for
example, in a C being converted to a U.
Mismatches of the normal bases because of a
failure of proofreading during DNA replication.
Common example: incorporation of the
pyrimidine U (normally found only in RNA)
instead of T.

Other damages
Breaks in the backbone.
Can be limited to one of the two strands (a singlestranded break, SSB) or
on both strands (a double-stranded break (DSB).

Ionizing radiation is a frequent cause, but some

chemicals produce breaks as well.
Crosslinks Covalent linkages can be formed
between bases
on the same DNA strand ("intrastrand") or
on the opposite strand ("interstrand").

Several chemotherapeutic drugs used against

cancers crosslink DNA [Link].

Where
Mitochondria more than nuclear DNA
Intracellular source is Mitochondrial
electron transport may generate radicals
ROS
Prevented by low calorie intake and free
radical inhibitors
Fe and Cu are associated with ROS

Membranes
Mitochondria
Enzymes
Chromosomes
DNA

Scientific American, Dec. 1992

Repairing Damaged Bases

Damaged or inappropriate bases can be repaired by
several mechanisms:
Direct chemical reversal of the damage
Excision Repair, in which the damaged base or bases are
removed and then replaced with the correct ones in a
localized burst of DNA synthesis. There are three
modes of excision repair, each of which employs
specialized sets of enzymes.
Base Excision Repair (BER)
Nucleotide Excision Repair (NER)
Mismatch Repair (MMR)

Cancer: What can we do about it?

Cancer Chemotherapy
The hallmark of all cancers is continuous cell division.
Each division requires both
the replication of the cell's DNA (in S phase) and
transcription and translation of many genes needed for
continued growth.
So, any chemical that damages DNA has the potential
to inhibit the spread of a cancer.
Many (but not all) drugs used for cancer therapy do
their work by damaging DNA.

What is the problem?

Sadly,
the cancer patient has many other cell types that are
also proliferating rapidly, e.g., cells of the
intestinal endothelium
bone marrow
hair follicles
and anticancer drugs also damage these producing
many of the unpleasant side effects of "chemo".
Agents that damage DNA are themselves carcinogenic,
and chemotherapy poses a significant risk of creating a
new cancer, often a leukemia.

Cancer Prevention
1) Many cancers are due to environmental exposures.
Limit the exposure
limit cancer.
(Smoking, UV-light, chemicals in preserved meat, etc.)
2) Viruses contribute to cancer formation.
Prevent viral infection

limit cancers

example: Human papilloma viruses (HPV)

common virus responsible for > 70% of cervical cancer
effective vaccine developed (2005)
politicized issue over its use

Early Diagnosis is
Important in Cancer Treatment
Goal: Identify tumor
before transition
to malignant stage

Traditional Treatments for

Tumors
Logic: Treatment must be targeted against
difference between normal and tumor cells
A) Fast Growing
B) Loss of Cell Cycle Control

What is our nature doing?

Cells Use Checkpoints to Fix

DNA Damage Before Cell Division
Key Point:
Cell cycle arrest
provides time to fix
damage before proceeding

Damaged
DNA
Stops
cell cycle

p53 = (Checkpoint protein

& Tumor suppresor)

Normal Cells with Extensive

Damage Commit Suicide
(Apoptosis)

Damaged DNA

p53

Too much
damage
Trigger cell death

Antioxidant Enzymes
The antioxidant enzymes are proteins with
antioxidant properties. There are three known classes
of antioxidant enzymes:
Superoxide dismutases (SOD)
Catalases (CAT)
Peroxidases (Glutathione Peroxidase, GP)
There are many forms of each class of protein. In
general, cancer cells have low levels of these enzymes,
when compared to an appropriate normal cell control.

Primary Antioxidant Enzyme

System

http://lpi.oregonstate.edu/f-w97/reactive.html

Another important concept in

cancer is Angiogenesis

What is Angiogenesis?
Angiogenesis is the
formation of new
capillary blood vessels
from quiescent
vascular endothelium.

Formation of new blood vessels

When EC and smooth
muscle cells grow in
response to specific signals
However, blood vessels do
not grow in healthy adult

The process of Angiogenesis

Local tissues require
additional O2 and nutrients
to sustain their growth.
Hypoxia results in the
upregulation of genes for
VEGF and enzymes that
trigger the process.

The process of Angiogenesis

For new blood
vessels to sprout
require that the
surrounding area be
broken down by
metalloproteinase
enzymes that carve
out the pathway for
vessel formation.

The process of Angiogenesis

This creates local
inflammation resulting
in migration of
macrophages and
inflammatory cells to
the site. Macrophages
and monocytes release
growth factors such as
FGF that synergize with
the VEGF activated by
hypoxia.

The process of Angiogenesis

The growth factors act
upon receptors located on
the surface of endothelial
cells

The process of Angiogenesis

The growth factors
message is in turn carried
into the cell by tyrosine
kinases which turn on the
machinery to initiate

ANGIOGENESIS

Types of Angiogenesis
Physiological Pathological

Physiological Angiogenesis
A developing child in a
mothers womb must create the
vast network of arteries, veins,
and capillaries that are found in
the human body. A process
called vasculogenesis creates
the primary network of vascular
endothelial cells that will
become major blood vessels.
Later on, angiogenesis remodels
this network into the small new
blood vessels or capillaries that
complete the childs circulatory
system.

Physiological Angiogenesis

The Bodys Control of Angiogenesis

The healthy body controls angiogenesis
through a series of "on" and "off" switches:
The main "on" switches are known as
angiogenesis-stimulating growth factors
The main "off switches" are known as
angiogenesis inhibitors

Pathological Angiogenesis

How can food help?

Omega-3 fatty acids and cancer therapy

W. Elaine Hardman, Ph.D.
Department of Biochemistry and Microbiology
Marshall University School of Medicine
Huntington, West Virginia

Outline
What are omega 3 fatty acids?
Pre-clinical evidence for benefit of n-3 fatty acids during cancer
therapy
Potential mechanisms for therapeutic benefit of n-3 fatty acids
Clinical evidence for benefit of n-3 fatty acids during cancer
therapy

Major fat types

O

Saturated fat
Stearic
18:0

17 15 13 11

Monounsaturated fat
18 16
Oleic
(OA)
18:1n-9

OH

O
12

OH

Polyunsaturated fats
O

Linoleic
(LA)
18:2n-6

Linolenic 18
(LNA)
18:3n-3

1312

OH

18

O
15

12

OH

Pre-clinical evidence for benefit of omega-3 fatty

acids during cancer therapy

Supplementing the diet with omega-3 fatty acids may

suppress the growth of existing cancers and may prevent or
slow metastasis
Omega 3 fatty acids may increase the efficacy of chemo- or
radiation therapy

Hormone responsive tumors such as:

breast, prostate and colon cancers
seem especially sensitive to omega 3 fatty acids.

However, in animal models, lung cancer growth has

been also slowed by omega 3 fatty acids.

Mean tumor size (mm )

MDA-MB 231 growth after initiation of DOX

900

Corn oil diet

800
700
600
500
400
300

Omega 3 diet

200
100
0
0

10

15

20

25

30

Number days after initiation of DOX

Growth rate mm3/day SEM
24.2 1.1
N-3
CO
3.3 0.6
N-3dox
COdox

35

5.1 0.4
-0.1