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目的：探究二十碳五烯酸(Eicosa Pentaenoic Acid. EPA)对SMMC-7721 人肝癌细胞的凋亡、端粒逆转录酶hTERT 的调控作用及端粒酶表达活性的影响。方法：体外培养SMMC-7721 人肝癌细胞，用不同浓度的EPA(0 uM、25 uM、50 uM、100 uM、200uM)作用于SMMC-7721 肝癌细胞（24 h、48 h、72 h）后，显微镜下观察其形态学变化；应用MTT 法检测SMMC-7721 肝癌细胞细胞增殖变化情况；Western-blot 法检测hTERT、Bax、Bcl-2 蛋白表达水平变化；Real Time-PCR 检测hTERTmRNA 的表达变化； ELISA 法检测SMMC-7721 肝癌细胞端粒酶活性的表达水平。结果：EPA可诱导肝癌细胞SMMC-7721 发生细胞凋亡，具有明显的时间计量依赖关系。在此过程中Bcl-2蛋白表达的降低和Bax蛋白表达上调，同时端粒酶逆转录酶hTERT 蛋白及其mRNA的表达水平和端粒酶活性均明显降低。结论：抑制端粒酶逆转录酶基因(hTERTmRNA)表达而抑制端粒酶的活性、诱导癌细胞凋亡，可能是EPA 的抗癌作用机制之一。

英文摘要:

Objective:To study the influence of Eicosa Pentaenoic Acid (EPA) on the apoptotic, the regulation of hTERT and telomerase activity of human hepatocarcinoma SMMC-7721 cell.Methods:Human hepatocarcinoma SMMC-7721 cell was cultured in vitro and treated with different concentrations of EPA (0 uM, 25 uM, 50 uM, 100 uM, 200 uM) for 24 h, 48 h, 72 h. The morphological changes of SMMC-7721 hepatoma cell were detected by microscope. MTT assay was explored to detect the proliferation of SMMC-7721 cells; The expression levels of hTERT, Bcl-2 and Bax protein were detected by Western blotting; The expression levels of SMMC-7721 cell hTERT mRNA were analyzed by Real time-PCR; ELISA was used to detect SMMC-7721 cell telomerase activity levels.Results:EPA induced SMMC-7721 cells apoptosis significantly, and the apoptosis rate was both in time-and dose-dependent manner. Western blotting showed that expression levels of hTERT and Bcl-2 protein was down-regulated and expression levels of Bax protein was up-regulated concurrently along with the apoptotic process, and the expression of hTERT mRNA as well as activity of telomerase decreased concurrently.Conclusion:EPA can restrain the activity of telomerase and induce death of SMMC-7721 cancer cells by inhibiting the expression of hTERT mRNA, it may be one of the anti-cancer mechanisms of EPA.

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