人胚肺成纤维细胞复制性衰老过程中端粒相关因子的表达*★

摘要

BACKGROUND: Telomere-associated proteins wil directly affect the function of telomeres, adjust the length of telomeric DNA, which are closely related with cellsenescence and carcinogenesis. OBJECTIVE: To find the key regulatory molecules in the cellsenescence process through observing the telomere-associated factor expression in normal cel replicative senescence process. METHODS: Based on established cel replicative senescence model, reverse transcription-PCR and western blot were used to detect the telomere-associated factor expression on the molecular and protein levels, including the telomere-associated factor human telomere binding protein 1, tankyrase 1, telomerase RNA, telomere protection protein 1 and P53 expressions in the human embryonic lung fibroblast replicative senescence. RESULTS AND CONCLUSION: The results showed that with the cellsenescence, transcription of human telomere binding protein 1 did not changed, while the protein expression of human telomere binding protein 1 was increased gradual y and then decreased rapidly; mRNA and protein expressions of telomere protection protein 1 did not changed; with the human embryonic lung fibroblast replicative senescence, expression of telomere protection protein 1 was decreased gradual y; with cellsenescence, telomerase RNA component showed an increasing trend; protein expression of P53 did not changed. Human telomere binding protein 1, telomere protection protein 1 and telomerase RNA play an important role in cellsenescence.%　　背景：端粒相关蛋白直接影响端粒的功能，调节端粒 DNA 的长度，与细胞的衰老和癌变密切相关。目的：通过观察正常细胞复制性衰老过程中端粒相关因子的变化来找寻细胞正常衰老过程中的关键调控分子。方法：在构建好的细胞复制性衰老模型基础上，利用 RT-PCR 与 western blot 方法在分子与蛋白水平检测端粒相关因子的表达变化，主要检测人胚肺成纤维细胞在复制性衰老过程中端粒相关因子人端粒结合蛋白1、端锚聚合酶1、端粒酶 RNA、端粒末端保护蛋白1以及 P53的表达情况。结果与结论：结果显示，随着细胞的衰老，人端粒结合蛋白1的转录水平未发生改变，而人端粒结合蛋白1的蛋白表达水平有逐渐升高而后快速降低的过程；端锚聚合酶1的 mRNA 水平及蛋白表达水平未发生变化；端粒末端保护蛋白1随着人胚肺成纤维细胞的衰老表达水平逐渐降低；端粒酶 RNA 组分随着细胞的衰老呈递增趋势；P53的蛋白表达未发生变化。综上认为，人端粒结合蛋白1、端粒末端保护蛋白1及端粒酶 RNA 在细胞的复制性衰老过程中起重要作用。