Antisense glutaminase inhibition decreases glutathione antioxidant capacity and increases apoptosis in Ehrlich ascitic tumour cells.
Glutamine is an essential amino acid in cancer cells and is required for the growth of many other cell types. Glutaminase activity is positively correlated with malignancy in tumours and with growth rate in normal cells. In the present work, Ehrlich ascites tumour cells, and their derivative, 0.28AS-2 cells, expressing antisense glutaminase mRNA, were assayed for apoptosis induced by methotrexate and hydrogen peroxide. It is shown that Ehrlich ascites tumour cells, expressing antisense mRNA for glutaminase, contain lower levels of glutathione than normal ascites cells. In addition, 0.28AS-2 cells contain a higher number of apoptotic cells and are more sensitive to both methotrexate and hydrogen peroxide toxicity than normal cells. Taken together, these results provide insights into the role of glutaminase in apoptosis by demonstrating that the expression of antisense mRNA for glutaminase alters apoptosis and glutathione antioxidant capacity.
Lora
Jorge
J
Departamento de Biología Molecular y Bioquímica, Laboratorio de Química de Proteínas, Facultad de Ciencias, Universidad de Málaga, Spain.
Alonso
Francisco J
FJ
Segura
Juan A
JA
Lobo
Carolina
C
Márquez
Javier
J
Matés
José M
JM
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D017209:Apoptosis; D002286:Carcinoma, Ehrlich Tumor / Q000235:genetics* / Q000473:pathology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival; D004247:DNA / Q000737:chemistry; D053938:DNA Fragmentation; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D005972:Glutaminase / Q000037:antagonists & inhibitors* / Q000235:genetics*; D005978:Glutathione / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D010718:Phosphatidylserines / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species; D013997:Time Factors
D000975:Antioxidants; D016376:Oligonucleotides, Antisense; D010718:Phosphatidylserines; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D004247:DNA; D006861:Hydrogen Peroxide; D005980:Glutathione Reductase; C505409:Casp3 protein, mouse; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D005972:Glutaminase; D005978:Glutathione; D008727:Methotrexate