M' Antonella, B' Francesca, G' Ludovica, B' Claudia

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Microtubule-interfering activity of parthenolide

• M. Antonella, B. Francesca, G. Ludovica, B.
  Claudia
• Chemico-Biological Interactions 149 (2004)165-173

Presented by..Apichai E-wan
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Cytoskeleton

• Cell structure proteins within the cytoplasm of
  eukaryotic cells.
• Cellular functions
• Maintenance of cellular shape
• Cellular movement
• Intracellular transport
• Cell division

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Cytoskeleton
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Microtubules

• Tubulin dimer, consisting of a-tubulin and
  ß-tubulin
• Maintenance cell shape
• Cell motility
• Chromosome movement in cell division
• Organelle movement

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Microtubule assembly
Protofilament assembly
Sheet assembly
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Microtuble elongation
Minus end
Plus end
ß
a
-

GTP
GDP
GTP
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Drugs that disrupt microtubules

• Colchicine
• Depolymerization
• Vinblastine and vincristine
• Depolymerization
• Taxol Placlitaxel
• Stabilize

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Parthenolide

• Sesquiterpene lactone
• Feverfew
• Anti-inflammatory
• Cytotoxicity

Tanacetum parthenium
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Materials Methods

• Microtubule assembly assay
• Fluorescence microscopy
• Cytotoxic assay

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Microtubule assembly assay
DMSO
1 µM P
Continuous abs. 350 nm at 37 ?C 30 min
Tubulin solution
10 µM P
P Parthenolide PTX Placlitaxel
10 µM PTX
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(No Transcript)
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Fluorescence microscopy
Microtubule staining
24 hrs.
Fresh medium with test cpn.
TEST CPN.
MCF-7 CELLS
Fix
Rinse
Primary Ab
Wash
Secondary Ab
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Fluorescence microscopy
Chromatin staining
24 hrs.
Fresh medium with test cpn.
TEST CPN.
MCF-7 CELLS
Fix
DAPI
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Fluorescence microscopy
Microtubule staining
Chromatin staining
WASH
AIR DRIED
FLUORESCENCE PATTERN
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Fluorescence microscopy
Microtubule staining
A
B
C
D
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Fluorescence microscopy
chromatin staining
A
B
C
D
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Cytotoxicity assay
24 hrs.
TEST CPN.
EXPO. CELLS
trypan blue
trypsinized
Microscopical determination
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Pathenolide
Pathenolide/Paclitaxel
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Cytotoxicity assay

• Cell growth
• Reduced MCF-7 cell growth
• Dose dependent
• Combined treatment
• 40 reduction of cell viability if administration
  alone
• Enhance cytotoxicity

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CONCLUSION
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CONCLUSION

• Sesquiterpene lactone
• Medicinal herb feverfew
• Cytotoxicity
• Increase sensitivity to Paclitaxel in breast
  cancer MCF-7 cells

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CONCLUSION
Parthenolide is a chemotherapeutic drug in
combination with other microtubule-interfering
agents.
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Thank you for your attention