Mechanistic aspects of the induction of apoptosis by lauryl gallate in the murine B-cell lymphoma line Wehi 231.
The effect of lauryl gallate (antioxidant E-312) has been studied on the mouse B-cell lymphoma line Wehi 231. This compound is able to inhibit protein tyrosine kinases (PTKs) in whole cells and in crude extracts with a better efficiency than other well-known PTK inhibitors such as herbimycin or genistein. Initial events triggered upon the incubation of cells with lauryl gallate in phosphate-buffered saline (up to 1 h) include the inhibition of tyrosine phosphorylation, discharge of the mitochondrial transmembrane potential, and induction of mRNA for Bcl-2. Long-term cultures in complete medium supplemented with fetal calf serum (up to 24 h) in the presence of this compound exhibit clear apoptotic features such as increase in phosphatidylserine in the cell surface, decrease in the functionality of mitochondria, cytochrome c release to the cytosol, activation of caspases, hypodiploidy, and oligonucleosomal breakdown of DNA. Comparison between Wehi cells overexpressing Bcl-2 (Wehi-bcl-2) with Wehi-neo cells shows a delay in the manifestations of the apoptotic signs, indicating that Bcl-2 has a partial protective effect on the apoptosis induced by lauryl gallate. The proapoptotic effect of lauryl gallate is not dependent on DNA or protein synthesis, is not blocked by the chelation of calcium, and is not reverted by N-acetylcysteine.
Roy
G
G
Servicio de Immunología, Hospital Ramon y Cajal, Madrid, Spain.
Lombardía
M
M
Palacios
C
C
Serrano
A
A
Cespón
C
C
Ortega
E
E
Eiras
P
P
Lujan
S
S
Revilla
Y
Y
Gonzalez-Porqué
P
P
D000111:Acetylcysteine / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D016227:Benzoquinones; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D003574:Cytochrome c Group / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004171:Diploidy; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005434:Flow Cytometry; D005520:Food Preservatives / Q000737:chemistry / Q000494:pharmacology; D005707:Gallic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism; D006540:Herbicides / Q000494:pharmacology; D007700:Kinetics; D047029:Lactams, Macrocyclic; D016393:Lymphoma, B-Cell / Q000737:chemistry / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000187:drug effects; D010718:Phosphatidylserines / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017828:Rifabutin / Q000031:analogs & derivatives; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
D000975:Antioxidants; D016227:Benzoquinones; D003574:Cytochrome c Group; D005520:Food Preservatives; D006540:Herbicides; D047029:Lactams, Macrocyclic; D010718:Phosphatidylserines; D019253:Proto-Oncogene Proteins c-bcl-2; D011809:Quinones; D012333:RNA, Messenger; D017828:Rifabutin; D012965:Sodium Chloride; C008259:lauryl gallate; D005707:Gallic Acid; C020754:herbimycin; D000255:Adenosine Triphosphate; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; D011505:Protein-Tyrosine Kinases; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D005978:Glutathione; D002118:Calcium; D000111:Acetylcysteine