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Drug‐sensitivity pattern analysis for study of functional relationship between gene products
Author(s) -
Hiwasa Takaki,
Shimada Hideaki,
Sakaida Tsukasa,
Kitagawa Motoo,
Kuroiwa Namiko,
Ochiai Takenori,
Takiguchi Masaki
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)00918-9
Subject(s) - transfection , complementary dna , gene , microbiology and biotechnology , biology , clone (java method) , gene product , gene expression , genetics
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 IκBα superrepressor and dominant‐negative IKKβ 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.