Identification of compounds that induce or repress telomerase activity

端粒酶活性受hTERT基因调控。将hTERT启动子（1.8kbp）克隆成报告载体，通过绿色荧光或荧光素酶活性监测hTERT转录。由于正常成纤维细胞的寿命有限，因此需要使正常成纤维细胞永生，以便使用监测细胞。用逆转录病毒感染hTERT基因，使细胞永生化。利用hTERT永生化正常成纤维细胞、NHF-hTERT和MRC-hTERT生成hTERT转录监测细胞。我们最终生成了nft - trp - luc和MRCT-TRPLuc。我们还构建了端粒酶阳性癌细胞衍生的监测细胞RKO-TRPLuc，用于筛选抗癌药物靶向hTERT转录。利用RKO-TRPluc细胞，我们筛选了抑制癌细胞中hTERT转录的天然化合物，鉴定出12种天然产物。其中2个样品对hTERT转录有较强的抑制作用。这两种化合物应该具有很强的抗癌活性。需要进一步的研究来阐明这些化合物的作用机制。

Telomerase activity was regulated by hTERT gene regulation. hTERT promoter (1.8kbp) was cloned into reporter vector which can monitoring hTERT transcription by green fluorescence or luciferase activity. Since normal fibroblasts have limited life span, normal fibroblasts need to be immortalized to using monitoring cells. hTERT gene was infected with retroviruses to immortalized cells. hTERT immortalized normal fibroblasts, NHF-hTERT and MRC-hTERT were used to generate hTERT transcription monitoring cells. We finally generated NHFT-TRP-Luc and MRCT-TRPLuc. We also constructed monitoring cell, RKO-TRPLuc, derived from telomerase positive cancer cells to screening anti-cancer drug targeted hTERT transcription. Using RKO-TRPluc cells, we screened natural compounds which suppress hTERT transcription in cancer cells, and 12 natural products were identified. Among them, two samples showed strong inhibition of hTERT transcription. These two compounds should be powerful anti-cancer activity. Further examination should be necessary to clarify the mechanism of these compounds.