咖啡因抑制由紫外光引起的hRRM1和hRRM2的受激转录.docVIP

  咖啡因抑制由紫外光引起的 hRRM1 和 hRRM2 的受激转录 5 10 15 20 25 30 35 40 45  摘要：紫外辐射是常见的外界刺激，可造成 DNA 损伤、皮肤癌等一系列危害。许多研究表 明服用含咖啡因的饮料可以降低与紫外辐射相关的皮肤癌的发生，但其发挥作用的机制尚不 十分明确。因为咖啡因可以抑制 ATM-ATR 激酶介导 DNA 损伤应答网络的活性，本文通过 检测受该应答网络调控的基因的转录来探索咖啡因降低与紫外辐射相关的皮肤癌的作用机 制。此研究以 HeLa 细胞作为人体细胞模型，在给予或者不给予咖啡因处理的条件下，使其 受到紫外光辐射。首先本文通过集落形成实验证实 HeLa 细胞在受到紫外光辐射后，咖啡因 促使其凋亡。然后通过对比两种不同条件下，受 ATM-ATR 调控的基因的转录变化，发现了 CHK1、p53、 E2F1 和 RNR 亚基基因 hRRM1、hRRM2 及 p53R2 转录受咖啡因的影响。本 文揭示在 HeLa 细胞受到紫外光刺激后，咖啡因使 CHK1 及 p53 基因的转录较显著增加，可 以促使 HeLa 细胞的细胞周期停止和程序性凋亡。同时咖啡因还使 RNR 亚基基因中 hRRM1 和 hRRM2 的受激转录受到明显抑制，从而抑制紫外光引起的 dNTP 库的增加。本文阐明咖 啡因通过抑制两种 RNR 亚基基因的受激转录阻碍 dNTP 库的增加，影响 DNA 损伤修复的 进行，从而不利于细胞在经紫外辐射后的生存，也降低了细胞带着未修复的 DNA 损伤继续 生存以致突变和癌变的可能。 关键词：生物化学与分子生物学；DNA 损伤应答；核苷酸还原酶；紫外光；咖啡因 中图分类号：Q523 Caffeine Suppresses the UV-induced Transcriptional Response of hRRM1 and hRRM2 in HeLa Cells ZENG Zhen, WANG Dan, WANG Zitian, ZHOU Zheng (College of Biology, Hunan University) Abstract: The most common environmental genotoxic agent, the ultraviolet light (UV), induces DNA lesions which induce skin carcinomas or mutations if unrepaired. It has been reported that taking caffeinated beverage significantly reduces the risk of UV-caused skin carcinomas, but the mechanism of caffeine suppressing cancer risk is unclear. Our data have demonstrated that caffeine reduces the UV survival of HeLa cells which was tested by the colony-forming assay. Since caffeine inhibits the activity of the ATM-ATR mediated DNA damage response (DDR) network, we speculated that the UV-induced transcriptional response of genes regulated by this network may influenced by caffeine. Our data display that the mRNA level of several genes, regulated by ATM-ATR DDR network, is affected by caffeine in HeLa cells following UV irradiation. Caffeine upregualtes the transcription of CHK1 and p53 but suppresses the transcription of hRRM1 and hRRM2, which code two ribonucleotide reductases that significantly affect dNTP pool in human cells. Since the repair of UV-caused DNA lesions requires in