In vitro impact of a whey protein isolate (WPI) and collagen hydrolysates (CHs) on B16F10 melanoma cells proliferation.
BACKGROUND
Porcine skin gelatine presented anti-tumoral effect on murine hepatoma cells (MH134), inducing programmed cell death (apoptosis). Whey proteins (mainly lactoferrin) have been investigated for cancer prevention and treatment.


OBJECTIVE
Investigation of the inhibitory capacity on melanoma cells (B16F10) proliferation and the influence on % distribution of cell cycle phases, in the presence of various concentrations of whey protein isolate (WPI), bovine collagen hydrolysate (BCH) or its fractions.


METHODS
The permeate fraction BCH-P1 (molecular mass, MM 2.5kDa) was further fractionated into five retentate fractions (R1-R5) by ultrafiltration membranes and into four fractions (F1-F4) by reverse phase chromatography. The permeate BCH-P1 and all its fractions were comparatively tested against a negative control (B16F10 cells+culture medium), and also against a positive control (B16F10+culture medium+WPI).


RESULTS
The inhibitory concentrations for 50% of B16F10 cells (IC(50)) ranged from 0.19 to 156.9 mg/mL for all these proteins evaluated. The most inhibitory fractions of the BCH hydrolysate were BCH-P1 and F1-F4 with IC(50) concentrations below 1mg/mL. Changes in cell cycle phases were characterized by a general decrease in the G2/M phase that emphasizes growth arrest, some increase in phase S (BCH-P1 and F4) but a strong increase in G0/G1 phase for BCH-P1 and F4. Caspase-3 expression increased significantly in all media containing F and R fractions, and also in the presence of BCH or WPI. Apoptosis was extremely high at low concentration (400 microg/mL) of the F1-F3 fractions.


CONCLUSIONS
It is suggested that a mechanism for tumorigenesis inhibition may involve the caspases cascade and apoptosis.
Castro
G A
GA
Food and Nutrition Department, Food Engineering Faculty, State University of Campinas (UNICAMP), Campinas, SP, Brazil. sgarb@fea.unicamp.br
Maria
D A
DA
Bouhallab
S
S
Sgarbieri
V C
VC
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D053148:Caspase 3 / Q000378:metabolism; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D003094:Collagen / Q000494:pharmacology*; D005780:Gelatin / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D008545:Melanoma / Q000378:metabolism*; D051379:Mice; D008894:Milk Proteins / Q000494:pharmacology*; D011492:Protein Hydrolysates / Q000494:pharmacology*; D012878:Skin Neoplasms / Q000378:metabolism*; D013552:Swine; D000067816:Whey Proteins
D000970:Antineoplastic Agents; D008894:Milk Proteins; D011492:Protein Hydrolysates; D000067816:Whey Proteins; D005780:Gelatin; D003094:Collagen; D053148:Caspase 3