N normal human lymphocytes. The majority of regular human cells have
N regular human lymphocytes. The majority of normal human cells have no detectable telomerase activity, even so, activity is normally detected in cancer cells. Thus, inhibiting telomerase activity and inducing apoptosis may possibly possess a selective impact on cancer cells. The aim from the present study was to investigate the inhibitory effects of telomerase activity by CAUE inside a NALM-6 cell PAK6 Compound culture technique. CAUE was shown to preferentially harm DNA NPY Y5 receptor Molecular Weight synthesis compared with RNA or protein synthesis. Additionally, telomerase activity was drastically suppressed and also the activity of human telomerase reverse transcriptase (hTERT), a subunit of telomerase, was decreased following treatment 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 would be the 1st to identify the cytotoxic mechanisms of CAUE in leukemia cells. Introduction Telomerase, a specialized ribonucleoprotein, plays an vital part in cell proliferation by defending against the problem of end-replication by adding TTAGGG repeats to telomeres (1). The majority of normal human cells have no detectable telomerase activity, nonetheless, activity is usually detected in cancer cells (2,3). The inhibition of telomerase causes a progressive and critical reduction of telomeres, top to a potent signal for the blockage of cell proliferation along with the induction of apoptosis (four). Targeting the inhibition of telomerase activity and also the induction of apoptosis may have a selective impact on cancer cells. Clinically, B-cell acute lymphoblastic leukemia is curable, even so, 50 of adults encounter treatment failure as a consequence of drug resistance plus the inability of older adults to tolerate the side-effects of therapy (5). As a result, it truly is desirable to develop novel anticancer drugs against B-cell leukemia, like those targeting the inhibition of telomerase activity, to prevent side-effects following chemotherapy. Our previous study reported that treatment with caffeic acid undecyl ester (CAUE), a novel caffeic acid derivative, decreased cell survival in human B-cell leukemia NALM-6 cells, but exhibited no important impact on the survival of typical lymphocytes. Moreover, the cytotoxic induction mechanisms of CAUE were shown to be involved in the intrinsic apoptotic pathway within a caspase-dependent manner (6). The present study focused around the inhibitory effects of telomerase activity by CAUE inside a NALM-6 cell culture method. Components and methods Supplies and cell culture. CAUE was ready as described previously (7). All other reagents, unless otherwise stated, were from the 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 manage (rabbit polyclonal; Cell Signaling Technologies, Inc., Danvers, MA, USA) were employed. Human B-cell leukemia NALM-6 cells had been supplied by the Cell Resource Center for Biomedical Research (Tohoku University, Sendai, Japan). Cell culture reagents had been obtained from Invitrogen Life Technologies (Carlsbad, CA, USA) and the cells had been routinely cultured applying normal solutions, as described previously (8,9). DNA, RNA and protein synthesis assays. The effect of CAUE on the synthesis of DNA.