Cells. The aim of the present study was to investigate the inhibitory effects of telomerase activity by CAUE inside a NALM-6 cell culture system. CAUE was shown to preferentially damage DNA synthesis compared with RNA or protein synthesis. Also, telomerase activity was considerably suppressed along with the activity of human telomerase reverse MMP-12 Inhibitor Molecular Weight transcriptase (hTERT), a subunit of telomerase, was decreased following therapy with CAUE, every single inside a concentration-dependent manner. These benefits indicated that the cytotoxic effects of CAUE are mediated by the inhibition of DNA synthesis and telomerase activity. The present study could be the initially to recognize the cytotoxic mechanisms of CAUE in leukemia cells. Introduction Telomerase, a specialized ribonucleoprotein, plays an critical role in cell proliferation by guarding against the issue of end-replication by adding TTAGGG repeats to telomeres (1). The majority of standard human cells have no detectable telomerase activity, having said that, activity is commonly detected in cancer cells (two,3). The inhibition of telomerase causes a progressive and critical reduction of telomeres, leading to a potent signal for the blockage of cell proliferation along with the induction of apoptosis (four). Targeting the inhibition of telomerase activity and the induction of apoptosis could have a selective impact on cancer cells. Clinically, B-cell acute lymphoblastic leukemia is curable, even so, 50 of adults knowledge therapy failure as a consequence of drug resistance along with the inability of older adults to tolerate the side-effects of therapy (5). For that reason, it can be desirable to develop novel anticancer drugs against B-cell leukemia, which includes these targeting the inhibition of telomerase activity, to stop side-effects following chemotherapy. Our earlier study reported that remedy with caffeic acid undecyl ester (CAUE), a novel caffeic acid β adrenergic receptor Agonist Purity & Documentation derivative, decreased cell survival in human B-cell leukemia NALM-6 cells, but exhibited no significant impact on the survival of normal lymphocytes. Moreover, the cytotoxic induction mechanisms of CAUE had been shown to become involved inside the intrinsic apoptotic pathway inside a caspase-dependent manner (six). The present study focused on the inhibitory effects of telomerase activity by CAUE within a NALM-6 cell culture program. Supplies and strategies Components and cell culture. CAUE was prepared as described previously (7). All other reagents, unless otherwise stated, had been of your highest grade accessible and bought from Sigma-Aldrich (St. Louis, MO, USA) or Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Antibodies against human telomerase reverse transcriptase (hTERT; rabbit polyclonal; Santa Cruz Biotechnology, Inc., Santa Cruz, CA USA) and -actin as the loading handle (rabbit polyclonal; Cell Signaling Technologies, Inc., Danvers, MA, USA) have been applied. Human B-cell leukemia NALM-6 cells had been supplied by the Cell Resource Center for Biomedical Investigation (Tohoku University, Sendai, Japan). Cell culture reagents have been obtained from Invitrogen Life Technologies (Carlsbad, CA, USA) and also the cells have been routinely cultured utilizing regular techniques, as described previously (8,9). DNA, RNA and protein synthesis assays. The impact of CAUE around the synthesis of DNA, RNA and protein was determined by incorporation from the radioactive precursors [3H]-thymidine, [3H]-uridine and [14C]-leucine (GE Healthcare, Amersham, UK). Briefly, 4×10 5 cells/ml had been cultured in 96-well round-bottom plates within a total volume of one hundred cu.