In vitro impact of a whey protein isolate (WPI) and collagen hydrolysates (CHs) on B16F10 melanoma cells proliferation

J Dermatol Sci. 2009 Oct;56(1):51-7. doi: 10.1016/j.jdermsci.2009.06.016. Epub 2009 Aug 19.


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.

Conclusion: It is suggested that a mechanism for tumorigenesis inhibition may involve the caspases cascade and apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Caspase 3 / metabolism
  • Cattle
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cell Cycle / drug effects
  • Cell Cycle / physiology
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Collagen / pharmacology*
  • Gelatin / pharmacology*
  • Inhibitory Concentration 50
  • Melanoma / metabolism*
  • Mice
  • Milk Proteins / pharmacology*
  • Protein Hydrolysates / pharmacology*
  • Skin Neoplasms / metabolism*
  • Swine
  • Whey Proteins


  • Antineoplastic Agents
  • Milk Proteins
  • Protein Hydrolysates
  • Whey Proteins
  • Gelatin
  • Collagen
  • Caspase 3