Cytotoxic mechanisms of inhibitor of RNA synthesis, 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS-106).
We investigated the molecular mechanisms of cell death induced by 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS-106: Figure 1), a potent inhibitor of RNA synthesis, using mouse mammary tumor FM3A cells and human fibrosarcoma HT1080 cells. ECyd induced the characteristics of apoptosis on these cells, such as morphological changes, DNA fragmentations and caspase-3-like protease activation. General caspases inhibitor (Z-Asp-CH2-DCB) inhibited cell death. Interestingly, we also found that ECyd induced rRNA fragmentation with the size of 3.2, 2.8 and 1.5 kb, and which might be caused by inhibition of RNA synthesis. rRNA fragmentation was mainly occurred in D8 domain of 28 S rRNA, and the end of 5'-terminal sequence of 1.5 kb fragment was C3220pC3221p or C3221pG3222p, that was identical to the recognition sequence of RNase L. Furthermore, the fragmentation patterns of rRNA digested with RNase L resembled that of ECyd treated cells in shape. These results indicate that antitumor mechanisms of ECyd are involved in activation of RNase L. rRNA fragmentation may be one of the death events as a result of inhibition of RNA synthesis and play an important role in the antitumor activity of ECyd.
Yokogawa
T
T
Faculty of Pharmaceutical Science, Okayama University, Tsushima, Okayama 700-8530, Japan.
Takenaka
K
K
Ogawa
D
D
Futagami
M
M
Matsuda
A
A
Fukushima
M
M
Kitade
Y
Y
Wataya
Y
Y
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D003562:Cytidine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004722:Endoribonucleases / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D051379:Mice; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D012334:RNA, Neoplasm / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism; D014407:Tumor Cells, Cultured
C103084:1-(3-C-ethynylribopentofuranosyl)cytosine; D000970:Antineoplastic Agents; D019384:Nucleic Acid Synthesis Inhibitors; D012334:RNA, Neoplasm; D012335:RNA, Ribosomal; D003562:Cytidine; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease