N regular human lymphocytes. The majority of typical human cells have
N standard human lymphocytes. The majority of normal human cells have no detectable telomerase activity, nevertheless, activity is typically detected in cancer cells. Hence, inhibiting telomerase activity and inducing apoptosis may possibly have a selective effect on cancer cells. The aim with the present study was to investigate the inhibitory effects of telomerase activity by CAUE in a NALM-6 cell culture program. CAUE was shown to preferentially damage DNA synthesis compared with RNA or protein synthesis. Additionally, telomerase activity was drastically suppressed plus the activity of human telomerase reverse transcriptase (hTERT), a subunit of telomerase, was decreased following 5-HT6 Receptor Modulator drug treatment with CAUE, every single in 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 determine the cytotoxic mechanisms of CAUE in leukemia cells. Introduction Telomerase, a specialized ribonucleoprotein, plays an vital RelA/p65 drug function in cell proliferation by safeguarding against the issue of end-replication by adding TTAGGG repeats to telomeres (1). The majority of normal human cells have no detectable telomerase activity, having said that, activity is typically detected in cancer cells (two,three). The inhibition of telomerase causes a progressive and crucial reduction of telomeres, major to a potent signal for the blockage of cell proliferation plus the induction of apoptosis (4). Targeting the inhibition of telomerase activity plus the induction of apoptosis may perhaps have a selective effect on cancer cells. Clinically, B-cell acute lymphoblastic leukemia is curable, on the other hand, 50 of adults experience remedy failure as a consequence of drug resistance as well as the inability of older adults to tolerate the side-effects of therapy (5). For that reason, it really is 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 preceding study reported that treatment with caffeic acid undecyl ester (CAUE), a novel caffeic acid derivative, lowered 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 were shown to become involved within the intrinsic apoptotic pathway in a caspase-dependent manner (six). The present study focused around the inhibitory effects of telomerase activity by CAUE in a NALM-6 cell culture program. Materials and approaches Components and cell culture. CAUE was ready as described previously (7). All other reagents, unless otherwise stated, had been in the highest grade available and purchased 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 because the loading handle (rabbit polyclonal; Cell Signaling Technologies, Inc., Danvers, MA, USA) were utilized. Human B-cell leukemia NALM-6 cells were supplied by the Cell Resource Center for Biomedical Study (Tohoku University, Sendai, Japan). Cell culture reagents had been obtained from Invitrogen Life Technologies (Carlsbad, CA, USA) plus the cells have been routinely cultured using common solutions, as described previously (eight,9). DNA, RNA and protein synthesis assays. The effect of CAUE on the synthesis of DNA.
