Validation of a polymerase chain reaction aided transcript titration assay (PATTY) for topoisomerase II in lung cancer samples

• Published in: Anticancer research Vol. 20; no. 5B; p. 3549
• Main Authors: Dingemans, A M, Van Ark-Otte, J, Smit, E F, Postmus, P E, Giaccone, G
• Format: Journal Article
• Language: English
• Published: Greece 01.09.2000
• Subjects: Antigens, Neoplasm; Autoradiography; Carcinoma, Non-Small-Cell Lung - enzymology; Carcinoma, Non-Small-Cell Lung - genetics; Carcinoma, Small Cell - enzymology; Carcinoma, Small Cell - genetics; DNA Topoisomerases, Type II - biosynthesis; DNA Topoisomerases, Type II - genetics; DNA-Binding Proteins; Humans; Isoenzymes - biosynthesis; Isoenzymes - genetics; Lung Neoplasms - enzymology; Lung Neoplasms - genetics; Polymerase Chain Reaction - methods; Reproducibility of Results; Ribonucleases; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Titrimetry; Tumor Cells, Cultured
• ISSN: 0250-7005

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.