Cathepsin L antisense oligonucleotides in a human osteosarcoma cell line: effects on the invasive phenotype.
Alterations in cathepsin L expression and trafficking have been associated with the progression and metastasis of several tumor entities. In the present study, we examined the effects of various cathepsin L antisense (as) phosphorothioate oligonucleotides on both the expression of cathepsin L and the invasive potential of the human osteosarcoma cell line MNNG/HOS. Seven oligonucleotides of 20-bp length each and one random control oligonucleotide were chosen to block cathepsin L expression. Northern blot analysis demonstrated a significant reduction in cathepsin L mRNA expression by the six antisense oligonucleotides at a concentration of 10 microM. Cathepsin L protein expression was reduced significantly (50-85%) by the antisense oligonucleotides, as compared with the controls. Adhesion to matrices of collagen I and matrigel was not affected. In in vitro motility and invasion assays performed in uncoated and precoated transwell chambers, the ability of cells to migrate through the filters was inhibited by 35-75% using antisense oligonucleotides. The random control did not show any inhibitory effect. These data demonstrate that in MNNG/HOS cells cathepsin L influences cellular malignancy by promoting migration and basement membrane degradation.
Krueger
S
S
Department of Pathology, Otto-von-Guericke University Magdeburg, Leipziger Strasse 44, 39120 Magdeburg, Germany. sabine.krueger@medizin.uni-magdeburg.de
Kellner
U
U
Buehling
F
F
Roessner
A
A
D001485:Basement Membrane / Q000378:metabolism; D015152:Blotting, Northern; D002401:Cathepsin B / Q000096:biosynthesis; D056646:Cathepsin F; D056657:Cathepsin K; D056668:Cathepsin L; D002403:Cathepsins / Q000096:biosynthesis / Q000235:genetics*; D002448:Cell Adhesion; D002455:Cell Division; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry; D003546:Cysteine Endopeptidases; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007797:Laminin / Q000737:chemistry; D009361:Neoplasm Invasiveness; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012516:Osteosarcoma / Q000235:genetics* / Q000628:therapy*; D010641:Phenotype; D011509:Proteoglycans / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D007797:Laminin; D016376:Oligonucleotides, Antisense; D011509:Proteoglycans; D012333:RNA, Messenger; D013778:Tetrazolium Salts; D013844:Thiazoles; C059485:matrigel; D003094:Collagen; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534281:CTSL protein, human; D056668:Cathepsin L; C534130:CTSK protein, human; D056657:Cathepsin K; C534090:CTSF protein, human; D056646:Cathepsin F; C022616:thiazolyl blue