Dietary whey proteins

and oxidative stress in

the prostate
Kyle D. Kent. J. Bomser and W. J. Harper
Department of Food Science and Technology
The Ohio State University
 The prostate
• Prostate cancer is the 2nd leading cause of
cancer death in men
– Oxidative stress is a major contributor
– Diets rich in antioxidants may prevent cancer
development
Cancer development

mutation

glutathione oxidative stress

 What is glutathione (GSH)?
• The major cellular antioxidant

• Composed of 3 amino acids

– glutamate, cysteine, glycine
• GSH synthesis is limited by cysteine availability
• GSH protects against oxidant-induced cell damage
 Why Whey?
• Cystine-rich protein source
• Whey proteins
– α-lactalbumin (4 cystine per molecule)
– β-lactoglobulin (2 cystine per molecule)
– bovine serum albumin (17 cystine per
molecule)
 Hypothesis
Digested whey protein isolate can elevate
intracellular GSH and protect against
oxidant-induced cell death in
human prostate cells
The objectives of the study were:
1. To determine if digested whey protein
isolate (WPI) can elevate GSH in human
prostate epithelial cells
2. To determine if digested WPI protects
against oxidant-induced cell death in
human prostate epithelial cells
 SDS-PAGE of WPI
206 kDa
91 kDa

51 kDa

35 kDa

28 kDa

20 kDa

7 kDa

A B C D E
(A) undigested WPI (B) digested WPI (C) α-lactalbumin
(D) β-lactoglobulin (E) bovine serum albumin
 Objective #1:

Can digested WPI

elevate intracellular
GSH in human prostate
cells?
 Materials
• Human prostate epithelial cells (RWPE-1)
• Digested whey protein isolate (d-WPI)
• Digested sodium caseinate (d-casein)
• N-acetylcysteine (NAC)
• Buthionine sulfoximine (BSO)
 Concentration and time dependent
GSH elevation
5 6
(A) * (B) *
4 * 5 * *
reduced glutathione
(nmol/10 6 cells)

reduced glutathione
(nmol/10 6 cells)
3
3
2
2
1 1

0 0
0 125 250 500 0 12 24 48
WPI (µ g/mL)
d-WPI time (hrs)

* significant difference from the control (p<0.05).

Cysteine availability and
GSH concentration
6

5
(A) *
*
reduced glutathione
(nmol/106 cells)

0
control d-casein d-WPI NAC

* significant difference from the control (p<0.05).

Inhibition of GSH synthesis by BSO
5
*
4
reduced glutathione
(nmol/10 cells)

3
6

2
*
1 *
0
BSO+
control BSO d-WPI
d-WPI
* significant difference from the control (p<0.05).
 Objective #2:

Can digested WPI

protect human prostate
cells against oxidant-
induced cell death?
 Materials
• Human prostate epithelial cells (RWPE-1)
• Digested whey protein isolate (d-WPI)
– 500 μg/mL, 24 hrs
• Buthionine sulfoximine (BSO)
– 500 μM, 24 hrs
• t-butyl hydroperoxide (TBHP)
– 0-1000 μM, 24 hrs
Protection vs. TBHP-induced
cell death
140
control
120 d-WPI
BSO+d-WPI
cell proliferation

100
(% control)

80
60
*
40 *
20
0
0 20 40 60 80 100 120 140
TBHP (µ M)
*Significant difference from the control at each
concentration of TBHP (p<0.05).
 Conclusions
• Digested WPI elevates intracellular GSH of
human prostate cells
• Elevation of intracellular GSH by de novo
synthesis
• Digested WPI also protects against oxidant-
induced cell death
 Acknowledgments
• Joshua A. Bomser
• W. James Harper
• Institute of Food Technologists
• The Ohio State University Department of
Food Science & Technology