Drug-sensitivity pattern analysis for study of functional relationship between gene products.
We have developed a method that we call 'drug-sensitivity pattern analysis', or DSPA, for analysis of protein function. Cells are transfected with cDNA of the test molecule, followed by analysis of the sensitivity of the transfected cells to multiple growth-inhibitory drugs. If two cDNA products have similar functions, their transfected cells should show similar drug-sensitivity patterns. The cDNAs of some signaling molecules were transfected into NIH3T3 or Ha-ras-transformed NIH3T3 (ras-NIH) cells and stable transfectants, which expressed high amounts of the gene product, were isolated. Chemosensitivity of the transfected clone was compared with the parental cells by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide method using more than 40 drugs. The chemosensitivity changes caused by the transfected gene were calculated and expressed numerically as 'drug chemosensitivity index' (DCI). When the DCI values were analyzed by regression analysis, a significant positive relationship between IkappaBalpha superrepressor and dominant-negative IKKbeta and an inverse relationship between p53 and Mdm2 were consistent with previous reports. Thus, the DSPA method is useful for identifying functional similarities between gene products.
Hiwasa
Takaki
T
Department of Biochemistry, Graduate School of Medicine, Chiba University, Inohana 1-8-1, Chuo-ku, 260-8670 Chiba, Japan. hiwasa@med.m.chiba-u.ac.jp
Shimada
Hideaki
H
Sakaida
Tsukasa
T
Kitagawa
Motoo
M
Kuroiwa
Namiko
N
Ochiai
Takenori
T
Takiguchi
Masaki
M
D016475:3T3 Cells; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D004351:Drug Resistance; D051550:I-kappa B Kinase; D020672:I-kappa B Proteins / Q000235:genetics / Q000502:physiology; D066298:In Vitro Techniques; D051379:Mice; D000072000:NF-KappaB Inhibitor alpha; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology; D011506:Proteins / Q000235:genetics* / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000502:physiology
D018076:DNA, Complementary; D020672:I-kappa B Proteins; C000606205:Nfkbia protein, mouse; D011506:Proteins; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D016159:Tumor Suppressor Protein p53; D000072000:NF-KappaB Inhibitor alpha; D017346:Protein Serine-Threonine Kinases; C496564:Chuk protein, mouse; D051550:I-kappa B Kinase; C496562:Ikbkb protein, mouse; C496566:Ikbke protein, mouse; C022616:thiazolyl blue