A new mechanism of 6-((2-(dimethylamino)ethyl)amino)-3-hydroxy-7H-indeno(2,1-c)quinolin-7-one dihydrochloride (TAS-103) action discovered by target screening with drug-immobilized affinity beads.
6-((2-(Dimethylamino)ethyl)amino)-3-hydroxy-7H-indeno(2,1-c)-quinolin-7-one dihydrochloride (TAS-103) is a quinoline derivative that displays antitumor activity in murine and human tumor models. TAS-103 has been reported to be a potent topoisomerase II poison. However, other studies have indicated that cellular susceptibility to TAS-103 is not correlated with topoisomerase II expression. Because the direct target of TAS-103 remained unclear, we searched for a TAS-103 binding protein using high-performance affinity latex beads. We obtained a component of the signal recognition particle (SRP) as a TAS-103 binding protein. This component is a 54-kDa subunit (SRP54) of SRP, which mediates the proper delivery of secretory proteins in cells. We fractioned 293T cell lysates using gel-filtration chromatography and performed a coimmunoprecipitation assay using 293T cells expressing FLAG-tagged SRP54. The results revealed that TAS-103 disrupts SRP complex formation and reduces the amount of SRP14 and SRP19. TAS-103 treatment and RNAi-mediated knockdown of SRP54 or SRP14 promoted accumulation of the exogenously expressed chimeric protein interleukin-6-FLAG inside cells. In conclusion, we identified signal recognition particle as a target of TAS-103 by using affinity latex beads. This provides new insights into the mechanism underlying the effects of chemotherapies comprising TAS-103 and demonstrates the usefulness of the affinity beads.
Yoshida
Makoto
M
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa, 226-8501, Japan.
Kabe
Yasuaki
Y
Wada
Tadashi
T
Asai
Akira
A
Handa
Hiroshi
H
D000634:Aminoquinolines / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism*; D002457:Cell Extracts / Q000737:chemistry; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D006367:HeLa Cells; D006801:Humans; D007192:Indenes / Q000494:pharmacology*; D015850:Interleukin-6 / Q000378:metabolism; D007668:Kidney / Q000166:cytology; D008863:Microspheres; D008970:Molecular Weight; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D034622:RNA Interference; D018271:Signal Recognition Particle / Q000737:chemistry*; D013194:Staining and Labeling
D000634:Aminoquinolines; D002352:Carrier Proteins; D002457:Cell Extracts; D007192:Indenes; D015850:Interleukin-6; D004364:Pharmaceutical Preparations; D018271:Signal Recognition Particle; C106647:6-((2-(dimethylamino)ethyl)amino)-3-hydroxy-7H-indeno(2,1-c)quinolin-7-one