Levamisole induced apoptosis in cultured vascular endothelial cells.
To better understand the anticancer activity of Levamisole (LMS), which serves as an adjuvant in colon cancer therapy in combination with 5-Fluorouracil, this study analyses LMS' ability to induce apoptosis and growth arrest in cultured human micro- and macrovascular endothelial cells (ECs) and fibroblasts. Cells exposed (24 h) to Levamisole (range: 0.5 - 2 mmol l(-1)) alone or in combination with antioxidants (10 mmol l(-1) glutathione or 5 mmol l(-1) N-Acetylcysteine or 0.1 mmol l(-1) Tocopherol) were evaluated for apoptosis ((3)H-thymidine assays, in situ staining), mRNA/protein expression (Northern/Western blot), and proliferation ((3)H-thymidine incorporation). Levamisole dose-dependently increased apoptosis in ECs to 230% (HUVECs-human umbilical vein ECs), 525% (adult human venous ECs) and 600% (human uterine microvascular ECs) but not in fibroblasts compared to control cells (set as 100%). Levamisole increased in ECs integrin-dependent matrix adhesion, inhibited proliferation (-70%), reduced expression of survival factors such as clusterin (-30%), endothelin-1 (-43%), bcl-2 (-34%), endothelial NO-synthase (-32%) and pRb (Retinoblastoma protein: -89%), and increased that of growth arrest/death signals such as p21 (+73%) and bak (+50%). LMS (2 mmol l(-1))-induced apoptosis was inhibited by glutathione (-50%) and N-Acetylcysteine (-36%), which also counteracted reduction by Levamisole of pRb expression, suggesting reactive oxygen species and pRb play a role in these processes. The ability of LMS to selectively induce apoptosis and growth arrest in endothelial cells potentially hints at vascular targeting to contribute to Levamisole's anticancer activity.
Artwohl
M
M
Department of Internal Medicine III, Division of Endocrinology and Metabolism, University of Vienna, Vienna, Austria.
Hölzenbein
T
T
Wagner
L
L
Freudenthaler
A
A
Waldhäusl
W
W
Baumgartner-Parzer
S M
SM
D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002196:Capillaries / Q000166:cytology / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D051152:Clusterin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D019332:Endothelin-1 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D005978:Glutathione / Q000494:pharmacology; D006023:Glycoproteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007978:Levamisole / Q000494:pharmacology*; D008565:Membrane Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012501:Saphenous Vein / Q000166:cytology / Q000187:drug effects; D013936:Thymidine / Q000378:metabolism; D014316:Tritium; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects; D014599:Uterus / Q000098:blood supply; D050998:bcl-2 Homologous Antagonist-Killer Protein
D000975:Antioxidants; C494772:BAK1 protein, human; C493956:CDKN1A protein, human; C495242:CLU protein, human; D051152:Clusterin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D019332:Endothelin-1; D006023:Glycoproteins; D008565:Membrane Proteins; D018832:Molecular Chaperones; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D016160:Retinoblastoma Protein; D016159:Tumor Suppressor Protein p53; D050998:bcl-2 Homologous Antagonist-Killer Protein; D014316:Tritium; D007978:Levamisole; D005978:Glutathione; D013936:Thymidine; D000111:Acetylcysteine