title: Features of proliferation and in vitro drug resistance in central primitive neuro-ectodermal tumours.
abstract: The features of proliferation in brain tumours are related with clinical prognosis for several types of brain tumours, especially gliomas. For childhood central primitive neuro-ectodermal tumours (cPNET), including medulloblastoma, this relation has previously been unclear. The aim of this study is to investigate the relationship between proliferative features of cPNET and in vitro resistance for cytostatic drugs measured with the 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium-bromide (MTT) assay. Tumour material was obtained from 23 surgical specimens of cPNET. The expression of the proliferation markers Ki-67, proliferating cell nuclear antigen (PCNA) and cyclin D1 was determined with immunohistochemistry, while S-phase and DNA ploidy were analysed by flowcytometric analysis cell scan (FACS). The in vitro resistance for 10 cytostatic drugs was determined with the MTT assay. Drug resistance levels were available in 19 (83%) of the 23 samples with a complete profile of 10 cytostatic drugs tested in 14 samples. An excellent correlation in drug resistance scores was found between pharmacologically related drugs. The Ki-67 staining in 20 samples varied from 10 to 60% and from 30 to 100% for PCNA. Cyclin D1 staining was negative in 11 out of 18 samples. The S-phase in 16 samples ranged from 2 to 16%. Increased staining of Ki-67 was related with actinomycin D sensitivity (r -.603; P=0.022), while cells with a higher S-phase percentage were more resistant to ifosfamide (r.952; P<0.0001). In vitro drug resistance testing of central primitive neuro-ectodermal tumours (PNET) is feasible with the MTT assay. Ifosfamide resistance was related with increased Ki-67 and S-phase percentage of the tumour cells, while increased Ki-67 was also related with actinomycin D sensitivity. These findings suggest a cell cycle dependent activity of cytostatic drugs in vitro.
authors.affiliation: Department of Pediatrics, Subdivision Haemato-oncology, Vrije Universiteit Medical Centre, Amsterdam. n.schouten@vumc.nl
authors.forename: A Y N
authors.initials: AY
authors.lastname: Schouten van Meeteren
authors.forename: P
authors.initials: P
authors.lastname: van der Valk
authors.forename: H C
authors.initials: HC
authors.lastname: van der Linden
authors.forename: W J R
authors.initials: WJ
authors.lastname: van Ouwerkerk
authors.forename: A J F
authors.initials: AJ
authors.lastname: Broekhuizen
authors.forename: D R
authors.initials: DR
authors.lastname: Huismans
authors.forename: A H
authors.initials: AH
authors.lastname: Loonen
authors.forename: A J P
authors.initials: AJ
authors.lastname: Veerman
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D001932:Brain Neoplasms / Q000473:pathology* / Q000503:physiopathology*; D002455:Cell Division; D002469:Cell Separation; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D018242:Neuroectodermal Tumors, Primitive / Q000473:pathology* / Q000503:physiopathology*
chemical_list: D000970:Antineoplastic Agents