Molecular Radiobiology

Prepared by: Me
Interaction between radiation and tissue occurs at electron

level

Observable injury results from change in molecular level

Occurrence of molecular lesions is identified by effects on

macromolecules (direct) and effects on water (indirect)

Because the human body contains 80% water molecules,

radiation interaction with water is the principal radiation

interaction in the body

Ultimate damage occurs to the target molecule which

controls cellular metabolism and reproduction

When macromolecules are irradiated in vitro a considerable

radiation dose is required to produce a measurable effect

Irradiation in vivo demonstrates that macromolecules are

considerably more radiosensitive in their natural state

In vitro is irradiation outside of the cell or body

In vivo is irradiation within the body
 IRRADIATION OF MACROMOLECULES

When macromolecules
are irradiated in
solution in vitro, three
major effects occur:
main-chain scission,
cross-linking, and point
lesion
Main-chain scission

Breakage of the backbone of the long-chain macromolecule

Results to reduction of long to smaller macromolecules

Reduces size macromolecules (may still be macromolecular)

Reduces viscosity of solution

Measurements of viscosity determines the degree of main-

chain scission
Cross linking

Macromolecules produce spurs as a consequence of

irradiation
This spurs behave as though they have a sticky end and

attach to a neighbouring macromolecule or to another

segment of the same molecule
Increases the viscosity of the macromolecular solution
Point lesion

Disruption of single chemical bonds

Not detectable but can cause a minor modification in

the molecule
 Cell may malfunction and die

At low radiation doses, point lesions are considered to be

the cellular radiation damage that results in the late
radiation effects observed at the whole body
Synthesis of proteins and nucleic acids is critical to the

survival of the cell and to its reproduction

Radiation damage to any of these macromolecules may

result in cell death or late effects

Proteins are less radiosensitive than nucleic acids

 Metabolism consists of
catabolism (the reduction
of nutrient molecules for
energy) and anabolism
(the production of large
molecules for form and
function)
DNA is the most radiosensitive
molecule.
As the cell moves into S phase, the

ladder begins to open up in the

middle of each rung, much like a
zipper
Parent DNA is said to be replicated

into two duplicate DNA daughter

molecules

Half as much DNA is present in G1 as in G2

DNA damage can result to abnormal metabolic activity

Uncontrolled rapid proliferation of cells is the principal

characteristic of radiation induced malignant disease

With germ cells, response to radiation may not be observed

until the following generation, or even later

 RADIATION EFFECTS ON DNA

DNA is the most radiosensitive molecule

RADIATION RESPONSE OF DNA

A. Main-chain scission with

only one side rail severed

B. Main-chain scission with

both side rails severed

C. Main-chain scission and

subsequent cross-linking

D. Rung breakage causing a

separation of bases

E. Change in or loss of a base

Molecular lesions caused by damage

to the codon (triplet code) are called

point mutations, which may not be
reversible

Three principal observable effects

that may result from irradiation of

DNA are cell death, malignant disease,
and genetic damage
 Radiolysis of water
Irradiation of water represents the

principal radiation interaction in the

body
When water is irradiated, it dissociates

into other molecular products;

radiolysis of water
Ion pair can rejoin and become stable or the free electron

may attach to other water molecule

HOH+ and HOH- ions are unstable and can dissociate

into smaller molecules

Final result of radiolysis of water is the formation of an

ion pair, H+ and OH-, and two free radicals, H* and OH*.
 FREE RADICALS

Free radicals are highly

reactive, unstable, and
therefore exists with a lifetime
of less than 1 ms
Contains excess energy –
disrupt bonds and produce
point mutations
A free radical is an uncharged molecule that contains a single
unpaired electron in the outer shell
OH* can join with similar molecule to form hydrogen

peroxide
Hydrogen peroxide is poisonous to the cell –toxic agent
H* can interact with molecular oxygen to form

hydroperoxyl radical

Hydrogen peroxide (H2O2) and hydroperoxyl (HO*2) are

the principal damaging products after radiolysis of water

Hydrogen peroxide can also be
formed by interaction of two
hydroperoxyl

Organic molecules (RH) can become

reactive free radicals by irradiation

 In the presence of oxygen, another

type is possible

Free radicals are energetic molecules

because of their unique structure.

DIRECT AND INDIRECT EFFECTS

If the initial event occurs on the target molecule, the

effect of radiation is direct

The principal effect of radiation on human is indirect

Cellular Radiobiology
Prepared by: Me
Irradiation of a cell results to

possible changes in the target

molecule (DNA)
Response is either cellular

transformation or cell death

Cellular transformation –

stochastic effects
Cell death – deterministic effects
Target Theory
For a cell to die after radiation exposure, its target

molecule must be inactivated

 Originally used to represent cell lethality

Describe nonlethal radiation-induced cell abnormalities

DNA is the target molecule

The target is considered to be an area of the cell occupied

by the target molecule or by a sensitive site on the target

molecule

Hit – radiation interaction with the target

Hits occur through both direct and indirect effects
Cell Survival Kinetics

The lethal effects of radiation

are determined by observing
cell survival, not cell death
Single-target, single hit model applies to biologic

target, such as enzymes, viruses, and simple cells like

bacteria

Multitarget, single hit model applies to more

complicated biologic systems, such as humans
 Single-target, Single-hit Model

Poisson distribution – random

probability of a hit

Radiation interacts randomly with matter

 A hit is not simply an ionizing
event, but rather an ionization
that inactivates the target
molecule

D37 – radiation dose sufficient

enough to kill 63% of the cells’

population (37% survival)
Measure of radiosensitivity of the

cell
Low D37 indicates a highly

radiosensitive cell

High D37 reveals high

radioresistance

If there were no wasted hits (uniform interaction), D37 is the

dose that would be sufficient to kill 100% of the cells
Multitarget, single-hit model

Represents a threshold dose

Human cells have target number

ranging from 2 to 10
D0 : mean lethal dose - is a constant related to the

radiosensitivity of the cell (D37)

A large D0 indicates radioresistant cells, and a small D0 is
characteristic of radiosensitive cell

DQ : threshold dose

A measure of the width of the shoulder of the multitarget,
single-hit model and is related to the capacity of the cell to
recover from sublethal damage

A large DQ indicates that the cell can recover readily from

sublethal radiation damage
DOSES FOR VARIOUS EXPERIMENAL MAMMALIAN CELL LINES

Cell type D0 (rad) DQ (rad)

Mouse oocytes 91 62

Mouse skin 135 350

Human bone marrow 137 100

Human fibroblasts 150 160

Mouse 180 270

spermatogonia
Chinese hamster 200 210
ovary
Human lymphocytes 400 100
Recovery
The shoulder of the graph of the multitarget, single-hit

model shows that for mammalian cells, some damage must

be accumulated before the cell dies –sublethal damage
Wider shoulder means more sublethal damage can be

sustained and the higher the value of DQ

DQ is a measure of the capacity to accumulate sublethal

damage and the ability to recover from sublethal damage
 Cell cycle effects
Cell-cycle time or cell generation time – average time

from one mitosis to another

Most human cells – 24 hours

Cells in mitosis is are always most sensitive

G1 is the most time variable of cell phases

Human cells are most radiosensitive in M and most radioresistant

in late S
Changes in the genetic code of a germ cell can affect future
generations

Programmed cell death – apoptotic death

LET, RBE, and OER
At very high LET, cell survival kinetics follow the single-target,

single-hit model
With low LET radiation, the multitarget, single-hit model applies

Presence of oxygen maximizes the effect of low-LET radiation

When anoxic cells are exposed, a considerably higher dose is

required to produce a given effect

Irradiation of mammalian cells with high LET radiation follows

the single-target, single-hit model