PT - JOURNAL ARTICLE AU - Shwu-Bin Lin AU - Paul O. P. Ts'o AU - Shun-Kuo Sun AU - Kong-Bung Choo AU - Feng-Yuan Yang AU - Yun-Ping Lim AU - Hsiao-Lun Tsai AU - Lo-Chun Au TI - Inhibition of Thymidylate Synthase Activity by Antisense Oligodeoxynucleotide and Possible Role in Thymineless Treatment DP - 2001 Sep 01 TA - Molecular Pharmacology PG - 474--479 VI - 60 IP - 3 4099 - http://molpharm.aspetjournals.org/content/60/3/474.short 4100 - http://molpharm.aspetjournals.org/content/60/3/474.full SO - Mol Pharmacol2001 Sep 01; 60 AB - Thymidylate synthase (TS) is an important target for chemotherapeutic treatment of cancer. However, efficacy of TS-targeted anticancer drugs is limited by the development of drug resistance as a result of TS gene amplification. In this work, a phosphorothioated antisense oligonucleotide (ODN), designated ATS-2, was used to suppress cellular synthesis of TS. ATS-2 at 0.2 μM concentration was mixed with lipofectin in a charge ratio of 1:1 and was used to treat the human embryonic kidney (HEK) cell line. A reduction of TS mRNA and protein was achieved. Furthermore, a dose-dependent reduction of cumulative viable cells of up to 98% was observed. Flow cytometer analysis of cell cycle progression indicates that ATS-2-treated cells were arrested and went into apoptosis at the S phase, possibly because of thymidine shortage, suggesting that ATS-2 is specifically effective for dividing cells. When used in combination with the anticancer drug FdUrd, ATS-2 exerted a additive inhibitory effect on cellular proliferation. To elucidate the possible role of cellular thymidine kinase (TdR kinase) in ATS-2 treatment, a second cell line, HeLa, was used. Both HEK and HeLa have similar rates of cell division and ODN uptake. In contrast to HEK, which was shown to have very low levels of TdR kinase activity in [3H]thymidine incorporation experiments, [3H]thymidine incorporation in HeLa was 15-fold greater than that of HEK. We found that HeLa cells were sensitive to FdUrd but were rather resistant to ATS-2. On the contrary, HEK cells were sensitive to ATS-2 but insensitive to FdUrd. Effects of ATS-2 and FdUrd are, therefore, complementary in thymineless treatment too. The American Society for Pharmacology and Experimental Therapeutics