title: Colony formation of soft tissue sarcoma cells is inhibited by lipid-mediated antisense oligodeoxynucleotides targeting the human mdm2 oncogene.
abstract: More than one third of human soft tissue sarcoma (STS) have elevated levels of the MDM2 oncoprotein, resulting either from gene amplification or alternate mechanisms. MDM2 functions as a negative feedback regulator of the tumor suppressor p53. The aim of the present study was to investigate whether mdm2-antisense oligodeoxyribonucleotides (AS-ODNs) can influence the growth characteristics of two MDM2-overexpressing STS cell lines (US8-93, LMS6-93) where both have heterozygous p53 non-missense mutations. Cells were treated with lipofectamine-complexed mdm2 AS-ODNs complementary to a sequence of the mdm2 cDNA initiation site in comparison to sense control ODNs. After seeding and cultivation of a defined cell number the clonogenic survival was performed. The treatment of US8-93 cells with AS-ODNs, but not with sense ODNs, decreased the number of colonies up to > 80%. Western blot analysis demonstrated a significant decreasing of MDM2 protein level in AS-ODN transfected cells indicating an AS-specific inhibition of mdm2 transcription in US8-93 cells. Additionally, an increase of the G2/M population was found. In contrast, in the LMS6-93 cells treated with AS-ODNs only a decrease in clonogenic survival up to 26%, no change in MDM2 protein level and no cell cycle alterations were seen. All these factors taken together into consideration can be suggest that lipid-mediated mdm2 AS-ODNs could be as an effective therapeutic strategy for STS with an abnormal mdm2 overexpression.
authors.affiliation: Institute of Pathology, Faculty of Medicine, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Meye
authors.forename: P
authors.initials: P
authors.lastname: Würl
authors.forename: M
authors.initials: M
authors.lastname: Bache
authors.forename: F
authors.initials: F
authors.lastname: Bartel
authors.forename: U
authors.initials: U
authors.lastname: Grünbaum
authors.forename: J
authors.initials: J
authors.lastname: Mansa-ard
authors.forename: H
authors.initials: H
authors.lastname: Schmidt
authors.forename: H
authors.initials: H
authors.lastname: Taubert
mesh_terms: D002455:Cell Division / Q000235:genetics; D004337:Drug Carriers; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy; D006801:Humans; D008081:Liposomes; D009363:Neoplasm Proteins / Q000235:genetics; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000473:pathology; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000494:pharmacology / Q000627:therapeutic use; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051736:Proto-Oncogene Proteins c-mdm2; D012509:Sarcoma / Q000235:genetics* / Q000473:pathology* / Q000628:therapy; D014407:Tumor Cells, Cultured
chemical_list: D004337:Drug Carriers; D008081:Liposomes; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2