[关键词]
[摘要]
在人原发性肝癌中,转甲状腺素蛋白(transthyretin, TTR)基因转录严重受阻遏。在原发性肝癌患者的血浆中,TTR含量明显降低。因此,有必要从人血浆中提取TTR,观察TTR对人肝癌细胞生长的抑制作用。方法:先用40%饱和硫酸铵,再用5%酚预沉淀人血浆,采用DEAE-Cellulose离子交换层析提取TTR,所提蛋白经蛋白质印迹鉴定并于体外观察TTR对肝癌细胞生长的抑制作用。结果:从血浆中提取的TTR纯度为85%,经蛋白质印迹鉴定正确;体外实验初步证实,TTR对肝癌细胞的生长有明显抑制作用。结论: TTR对肝癌细胞的生长有明显抑制作用,这一性质为生物工程生产TTR,用于临床治疗,提供了可行性依据,从而在蛋白水平进一步证实TTR基因可能是与人肝癌相关的抑癌基因候选者。
[Key word]
[Abstract]
In primary hepatoma, the transcription of transthyretin (TTR) is seriously suppressed, and TTR decreased obviously in patients′ serum. It was necessary for us to purify TTR from human serum and observe the inhibitory effect of TTR on the growth of human hepatoma cells. Methods: The serum was precipitated in 40% saturated ammonium sulfate and 5% phenol, and the top layer was added to DEAE-Cellulose, then the protein was obtained. It was examined by Western blot. The inhibitory effect of TTR on the growth of human hepatoma cells was observed. Results: TTR was verified by Western blot and the purity of the TTR was at least 85%. It was confirmed that TTR inhibited evidently the growth of SMMC-7721 hepatoma cell. Conclusion: The evidently inhititory effect of TTR on SMMC-7721 hepatoma cell provided evidence for the feasibility of further genetic engineering production of TTR in future clinical therapy application. Italso supposed that TTR gene could be the candidate of the anti-oncogene related to human hepatoma
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[基金项目]
*本项目受国家攀登计划资助