5-FU vs 5-FU HeLa resistant

cells in terms of toxicity

BY: RAUNAK PRASAD
 INTRODUCTION TO 5-FU &
Chemotherapy drugs
● Chemotherapy drugs function by
disrupting the cell divisions- by
damaging DNA/RNA synthesis
● Different types of chemotherapy drugs-
alkylating, antimetabolite, plant
alkaloids
● 5-FU - antimetabolite
● A *heterocyclic aromatic organic
compound
● Which interferes with DNA replication
● Specific towards the S- Phase in cell
cycle (where copies of DNA and
duplication of centrosomes occurs)
Antimetabolites
● Antimetabolites are structurally
similar to the normal compounds that
exist within the cell.
● FUNCTION:
-> interferes the availability of normal
purines and pyrimidine nucleotide
precursors- by inhibiting the synthesis
of DNA/RNA
-> Cytotoxic effect is optimum at a
certain phase of the cell cycle- hence
called “cell cycle specific”.
-> exists in three kinds: folate
antagonist, purine antagonist, and
pyrimidine antagonist.
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 Mechanism of action

● 5-FU (inactive) -> 5-FdUMP-> forms a

complex with the thymidylate synthase->
inhibits dTMP production
● dTMP is needed for DNA replication and
repair, therefore its absence causes
cardiotoxicity.
● However, 5-FU can encounter resistance
when cells lose their ability to convert 5-
FU into its active form: 5-FdUMP… or
due to increase in levels of thymidylate
synthase.

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 Continued
ABC11/ MRP FAMILY
TS: THYMIDYLATE
SYNTHASE
● Catalyses the conversion of
dUMP to dTMP
● Inhibition causes imbalance of
deoxynucleotides and
increased levels of dUMP
● Thymidylate usually converts
uracil to thymidine by adding
a methyl group at the fifth
carbon of the pyrimidine
group.
● Formation of the ternary TS-
dUMP-CH2THF
● Reduced dTMP = reduced
dTTP = disturbances in the
nucleotide levels ->
disrupting DNA synthesis
and repair
Resistance Pathways
DPD- Dihydropyrimidine dehydrogenase
⩥ ABCC11 has shown to ● Mediates the conversion of 5-FU to DHFU- dihydrofluorouracil
be an active ● Becomes the rate limiting step of 5-FU catabolism
transporter of ● Up to 80% of administered 5-FU is broken down in the liver.
FdUMP, an active ● Found in the liver- which helps our body break down uracil and
metabolite for thymine
thymidine inhibition.
⩥ RT-PCR to compare the
expressions of ABCC1-
5, ABCC11, and
ABCC12 genes in the
5-FU resistant lung
cancer cell line.
⩥ Results- ABCC11 gene
found to be 2.5 fold
higher than the normal
parent cell line
Project Outline

● Conducted MTT sets to see the response of the

HeLa cells towards the effect of 5-Fluorouracil
in different concentrations
● Aim - Obtaining IC-50 Value under the right
conditions
● Concepts covered included-
thawing,trypsinizing, etc
● Making HeLa resistant cells
● 3 controls - vehicle and media
● Hypothesis - the data from the response of the
HeLa cells should be consistent with the HeLa
resistant cells in terms of cell viability
percentages.
● IC-50 Value found to be 2μg/mL.

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Results