title: Validation of a polymerase chain reaction aided transcript titration assay (PATTY) for topoisomerase II in lung cancer samples.
abstract: This report describes the validation of a polymerase chain reaction aided transcript titration assay (PATTY) for tumor samples. The results obtained with the PATTY were compared to those of RNase protection in a set of 7 human lung cancer cell lines and in 23 non-small cell lung cancer samples derived from resected patients. Whereas between PATTY and RNase protection assay a good correlation was observed in the cell lines (r = 0.74, p = 0.057), no correlation was observed within the tumor samples (r = 0.06, p = 0.78). This was also the case when only tumors with a high percentage of tumor cells (> 90%) were selected. Although PATTY is a valuable tool to measure mRNA expression in cell lines, our results caution the use of PATTY in human tumor samples without proper validation. The possible causes of these results are discussed.
authors.affiliation: Department Medical Oncology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands.
authors.forename: A M
authors.initials: AM
authors.lastname: Dingemans
authors.forename: J
authors.initials: J
authors.lastname: Van Ark-Otte
authors.forename: E F
authors.initials: EF
authors.lastname: Smit
authors.forename: P E
authors.initials: PE
authors.lastname: Postmus
authors.forename: G
authors.initials: G
authors.lastname: Giaccone
mesh_terms: D000951:Antigens, Neoplasm; D001345:Autoradiography; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology / Q000235:genetics; D018288:Carcinoma, Small Cell / Q000201:enzymology / Q000235:genetics; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis* / Q000235:genetics; D004268:DNA-Binding Proteins; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D008175:Lung Neoplasms / Q000201:enzymology* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D015203:Reproducibility of Results; D012260:Ribonucleases; D017186:Titrimetry; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D007527:Isoenzymes; D012333:RNA, Messenger; D012260:Ribonucleases; D004250:DNA Topoisomerases, Type II