Quantitative immunocytochemical assays of topoisomerase II in lung adenocarcinoma cell lines. Correlation to topoisomerase II alpha content and topoisomerase II catalytic activity.
The examination of topoisomerase II alpha content by Western blot analysis or topoisomerase II catalytic activity by decatenation of kDNA requires a large number of cells, but it is difficult to collect sufficient cells for these biochemical analyses from lung cancer patients by transbronchial brushing or aspiration. In this study, we explored the relationship between these biochemical analyses and topoisomerase II immunostaining in cytospin preparations of three lung adenocarcinoma cell lines. The levels of topoisomerase II alpha content were about 8.4 for A549, 2.9 for PC-3 and 1 for RERF-LC-MS, and the levels of topoisomerase II catalytic activity were about 4, 2, and 1, respectively. The percentages of strongly positive cells for topoisomerase II immunostaining were 60.9% for A549, 33.3% for PC-3, and 14.3% for RERF-LC-MS, and these were compatible with the levels of topoisomerase II alpha content or topoisomerase II catalytic activity. Our results indicate that topoisomerase II immunostaining can be utilized in place of biochemical analysis.
Yamazaki
K
K
First Department of Medicine, School of Medicine, Hokkaido University, Sapporo, Japan.
Isobe
H
H
Hanada
T
T
Sukoh
N
N
Ogura
S
S
Kawakami
Y
Y
D000230:Adenocarcinoma / Q000201:enzymology*; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm / Q000032:analysis*; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D015153:Blotting, Western; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002384:Catalysis; D004396:Coloring Agents; D004250:DNA Topoisomerases, Type II / Q000032:analysis* / Q000378:metabolism; D018105:DNA, Kinetoplast / Q000378:metabolism; D004268:DNA-Binding Proteins; D004317:Doxorubicin / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor; D005047:Etoposide / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000032:analysis*; D008175:Lung Neoplasms / Q000201:enzymology*; D014407:Tumor Cells, Cultured
D000903:Antibiotics, Antineoplastic; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D000972:Antineoplastic Agents, Phytogenic; D004396:Coloring Agents; D018105:DNA, Kinetoplast; D004268:DNA-Binding Proteins; D007527:Isoenzymes; D005047:Etoposide; D004317:Doxorubicin; D004250:DNA Topoisomerases, Type II