Aspirin Aspirin Protects Caco-2 Cells From Protects Caco-2 Cells From ApoptosisApoptosis After Serum Deprivation Through After Serum Deprivation Through

the Activation of a the Activation of a PI PI 3-Kinase/AKT/p213-Kinase/AKT/p21CipCip//WAFWAF11Pathway Pathway

P. Ricchi et al, P. Ricchi et al, Molecular Molecular PharmacologyPharmacology, , 6464, 2003, 2003

Ashim Malhotra November 11, 2003 Cell Signaling

PLA2PLA2

PLPL

COX 1/2COX 1/2

PGE2/ TXA2/ LKTsPGE2/ TXA2/ LKTs

Cell MembraneCell MembraneAA

The PI-3K PathwayThe PI-3K Pathway

InhibitsInhibits

Cox1/2Cox1/2

Arachidonate/ Prostaglandins/ Leukotriens Arachidonate/ Prostaglandins/ Leukotriens

Effect of Aspirin Treatment on Apoptosis Effect of Aspirin Treatment on Apoptosis Induced by Serum Deprivation in Caco-2 cellsInduced by Serum Deprivation in Caco-2 cells

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Fig. 01

Effect of 2 & 5 mM Aspirin Treatment on Effect of 2 & 5 mM Aspirin Treatment on Cell Cell Cycle DistributionCycle Distribution Observed After Caco-2 Observed After Caco-2

cells Serum Withdrawalcells Serum Withdrawal

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Fig. 02

Effect of 2 & 5 mM Aspirin Treatment on (Ser Effect of 2 & 5 mM Aspirin Treatment on (Ser 437) 437) Phosphorylated AKT & Total AKTPhosphorylated AKT & Total AKT

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Fig. 03

Effect of 2 & 5 mM AspirinTreatment on Effect of 2 & 5 mM AspirinTreatment on PI3-PI3-kinase Activitykinase Activity in 96-h Serum-Starved Cells in 96-h Serum-Starved Cells

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Fig. 04

Effect of 5 mM Aspirin Treatment on (Ser 437) Effect of 5 mM Aspirin Treatment on (Ser 437) Phosphorylated AKT & Total AKTPhosphorylated AKT & Total AKT & &

phosphorylated ERK1/2 & Total ERK2phosphorylated ERK1/2 & Total ERK2 ~O~~O~

Fig. 05

Effect of Effect of PD98059PD98059 & & LY294002LY294002 Treatments Alone & in Treatments Alone & in Combination with Aspirin on (Ser 437) Combination with Aspirin on (Ser 437) phosphorylated phosphorylated AKT & Total AKTAKT & Total AKT & & phosphorylated ERK1/2phosphorylated ERK1/2 & & Total Total

ERK2ERK2 Expression Expression

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Fig. 06

Effect of Effect of PD98059PD98059 & & LY294002LY294002 Alone or in Alone or in Combination with Aspirin on Apoptosis During Combination with Aspirin on Apoptosis During

Caco-2 StarvationCaco-2 Starvation

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Effect of Aspirin & Effect of Aspirin & Vp-16Vp-16 Treatments on the Treatments on the phosphorylation Status of phosphorylation Status of AKT ProteinAKT Protein

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Fig. 07

Effect of 40 µM Effect of 40 µM PD98059 PD98059 or 50 µM or 50 µM LY294002 LY294002 Treatments Alone or in combination with 2 & 5 mM Treatments Alone or in combination with 2 & 5 mM

Aspirin on Aspirin on p21p21CipCip//WAFWAF11 & -Tubulin & -Tubulin Expression in Serum- Expression in Serum-Starved CellsStarved Cells

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Fig. 08

Effect of Effect of PD98059 PD98059 & & LY94002 LY94002 Treatments Alone Treatments Alone or in combination with Aspirin on or in combination with Aspirin on Cell Cycle Cell Cycle

DistributionDistribution During Caco-2 Starvation During Caco-2 Starvation

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CCoonncclluussiioonnss

Aspirin treatment in Caco-2 cells interferes with serum deprivation induced apoptosis through PI –kinase & ERK kinase pathways.

PI-3 kinase mediates survival effect of aspirin.

Aspirin prevents an increase in the G2 cell cycle stage cells & dose dependently increases p21 expression.

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4 presentations!! 4 presentations!!

Phew!!Phew!!

AND now It is AND now It is

THE END!!!!!THE END!!!!!