Detection of differential gene expression in human osteoblastic cells by non-radioactive RNA arbitrarily primed PCR.
The aim of the present study was to detect differentially expressed genes in the human osteoblast-like osteosarcoma cell line SaOS-2 using non-radioactive RNA fingerprinting (RNA arbitrarily primed polymerase chain reaction, RAP-PCR). RNA was isolated at different time points from SaOS-2 cells grown with and without dexamethasone (DEX). By RAP-PCR we detected changes in band patterns of cells treated with DEX compared with untreated cells. PCR fragments further characterized and sequences from three of these gave perfect matches to the coding sequences of the human nucleophosmin gene B23, cDNA clone 4_c6 from P1 H25 and the human TRA1 gene, respectively. differential regulation of these genes in DEX-stimulated SaOS-2 cells could be demonstrated by RT-PCR.
Schnabel
M
M
Department of Traumatology, University Medical Center, University of Marburg, D-35043 Marburg, Germany.
Bortolussi
G
G
Fichtel
I
I
Kraus
A
A
Gotzen
L
L
Schlegel
J
J
D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D017931:DNA Primers; D003907:Dexamethasone / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012334:RNA, Neoplasm / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D018931:Antineoplastic Agents, Hormonal; D017931:DNA Primers; D012334:RNA, Neoplasm; D003907:Dexamethasone