Fenbendazole acts as a moderate

microtubule destabilizing agent and

causes cancer cell death by modulating
multiple cellular pathways

Muazzam Kazmi
Objective

1 The present work demonstrates a pleiotropic

effect of Fenbendazole (FZ) on cancer cells leading
to cell death.
2 FZ may have a potential therapeutic application.

2
Abstract

3
Abstract

1 Fenbendazole (FZ) demonstrates moderate affinity for

mammalian tubulin and exerts cytotoxicity to human cancer
cells at micromolar concentrations.

2 It caused mitochondrial translocation of p53 and effectively

inhibited glucose uptake, expression of GLUT transporters as
well as hexokinase (HK II) (a key glycolytic enzyme that
most cancer cells thrive on).

4
Abstract

1 FZ is a new microtubule interfering agent.

2 It displays anti-neoplastic activity and may be evaluated as

a potential therapeutic agent.
□ b/c of its effect on multiple cellular pathways leading to
effective elimination of cancer cells.

5
Introduction

6
Introduction

▣ The importance of microtubules in;

□ Cell division, motility, intracellular trafficking and their
role in modulating cellular shape according to the
environment
▣ has made them one of the most successful targets of
anticancer therapy.
▣ Agents that perturb the microtubule dynamics have been
widely used in cancer treatment.
▣ Considering the relative success of mitotic agents in the
treatment of cancer, microtubules may be termed as one of
the best cancer targets identified till now.
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Introduction

Microtubules:
▣ Microtubules are composed of a
single type of globular protein,
called tubulin.
▣ Tubulin is the major building block
of microtubules.
▣ This intracellular cylindrical
filamentous structure is present in
almost all eukaryotic cells.

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Introduction

▣ Tubulin is a heterodimer, which consists of:

□ α-tubulin and β-tubulin
□ both subunits have a molecular weight of 55 kDa and share
considerable homology.
□ α-tubulin and β-tubulin join together to form a dimer and put
them together.
▣ These dimers form long chains or polymers, these polymers will put
together into a sheets and then this sheet is going to be rolled up to
form a tube.

Sheet

tube 9
Introduction

▣ The diameter of microtubule is approximately 25 nm.

▣ One end of the microtubule is going to be anchored and is called
microtubule organizing center (MTOC)
▣ At the other end of microtubule dimers can be added very quickly to
make it longer or dimers could be taken off to make it shorter.
□ So it can become longer and shorter very quick
□ Microtubule are dynamic.
□ They change & change very quickly
▣ Its important for microtubule to become longer and shorter to fulfil its
functions.
Dimers can be added or remove

25 nm MTOC

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Introduction

▣ Microtubules are major components of the cytoskeleton.

▣ Play important roles in a variety of cellular processes
□ like intracellular trafficking, maintenance of cell shape and
structure, polarity, cell signaling, and mitosis.
▣ Microtubule targeting agents (MTAs) are being used
clinically for the treatment of multiple tumor types.
□ their effectiveness is often largely affected by drug resistance
mechanisms

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Introduction

▣ Microtubule targeting agents can be broadly classified into

two major classes.
□ Microtubule-destabilizing agents:
■ Inhibit microtubule polymerization.
■ Bind to the tubulin polymer and stabilize microtubules.
■ It includes several compounds such as the vinca alkaloids,
estramustine, colchicine and combretastatins
■ Used clinically or are under clinical investigation for cancer
treatment.
□ Microtubule-stabilizing agents:
■ Bind to the tubulin dimers and destabilize microtubules.
■ These agents include paclitaxel, docetaxel, epothilones, and
discodermolide.

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Introduction

The present data reveals:

□ FZ as a moderate microtubule targeting agent
causing mitotic arrest followed by cancer cell death.
□ Despite being a relatively mild microtubule
targeting agent, FZ possesses a unique ability to
induce p53 to a considerably high level.
□

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Introduction

▣ Fenbendazole (FZ) is a drug used typically not for

humans but for animals (fish, birds and amammals).
▣ It is labelled to kill worms such as roundworms,
hookworms, whipworms and some tapeworms.
▣ FZ is found under various brand names such as
Panacur or Safe-Guard

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Introduction

▣ Fenbendazole (FZ) is a broad-spectrum benzimidazole

anthelminthic approved for use in numerous animal species.
▣ Repurposing of veterinary drugs showing promising results
for human use can result in considerable time and cost
reduction required to develop new drugs.
▣ FZ is known to have a high safety margin and most species
tolerate it very well.
▣ It has very low degree of toxicity and high degree of safety
in experimental animals

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Introduction

▣ In this study, we show that FZ exhibits a moderate

microtubule depolymerizing activity towards human cancer
cells.
□ but possesses a potent antitumor effect.
▣ Our results indicate that:
□ FZ exerts its antitumor effect through the disruption of microtubule
dynamics.
□ p53 activation and the modulation of genes involved in multiple
cellular pathways.
▣ FZ treatment also resulted in reduced glucose uptake in
cancer cells due to down regulation of GLUT transporters and
key glycolytic enzymes.
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Introduction

▣ FZ could effectively inhibit glucose uptake in Human

non small cell lung carcinoma (NSCLC) cells.
□ suggesting that FZ induced cancer cell death is, in part,
facilitated by blocking glucose uptake of cancer cells.

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Results

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FZ destabilizes tubulin network in human
NSCLC cells

▣ Examine the effect of FZ on mammalian microtubule

network organization,
□ Human non small cell lung carcinoma (NSCLC) A549
cells were treated with 1 uM FZ for 24 h and
processed for immunofuorescence using α tubulin
antibody.
▣ Results showed that FZ treatment caused a partial
alteration of the microtubule network.

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FZ destabilizes tubulin network in human
NSCLC cells
The microtubule cage around the nucleus appeared to have
lost its intactness when compared with the control mock
treated cells.

A549 cells were treated with 1 uM FZ or 50 ng/ml colchicine for 24 h. Following treatment,
the cells were processed for immunofluorescence using anti α-tubulin primary and FITC
conjugated secondary antibodies. (Nuclei were counter stained with propidium iodide)

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FZ is not a P-gp substrate or inhibitor

To test whether FZ is a substrate

or inhibitor of P-gp:
▣ we investigated cancer cell
growth inhibition by FZ in the
presence of P-gp inhibitor
verapamil.
▣ The results showed that A549 and H460 cells were treated with 1 uM
inhibition of P-gp by verapamil FZ in the absence or presence of 10 uM
verapamil for 24 h
did not enhance the inhibitory
effect of FZ on cancer cell
proliferation.
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Conclusion

▣ Fenbendazole (FZ) is an new microtubule interfering

agent that displays anti neoplastic activity and may
be evaluated as a potential therapeutic agent because
of its effect on multiple cellular pathways leading to
effective elimination of cancer cells

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