| 刘建香,苏旭.重组质粒p-Egr-p16的构建及在SMMC-7721细胞中的辐射诱导表达[J].中华放射医学与防护杂志,2003,23(6):439-440.LIU Jianxiang,SU Xu.Construction of p-Egr-p16 plasmid and its expression induced by radiation in SMMC-7721 cells[J].Chin J Radiol Med Prot,2003,23(6):439-440 |
| 重组质粒p-Egr-p16的构建及在SMMC-7721细胞中的辐射诱导表达 |
| Construction of p-Egr-p16 plasmid and its expression induced by radiation in SMMC-7721 cells |
| 投稿时间：2003-05-25 |
| DOI： |
| 中文关键词: Egr-1基因 p16基因 p16 mRNA |
| 英文关键词:Egr-1 gene p16 gene p16 mRNA |
| 基金项目: |
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| 中文摘要: |
| 目的 构建重组质粒p-Egr-p16并探讨在SMMC-7721细胞中的辐射诱导表达及抗肝癌的作用。方法 用双酶切、粘端连接的方法构建了含有辐射诱导特性的早期反应因子Egr-1和p16的p-Egr-p16的质粒载体,以脂质体介导的方法,将重组载体导入人肝癌SMMC-7721细胞,采用RT-PCR的方法检测了不同剂量60Coγ射线照射转染后的人肝癌细胞p16基因转录水平的变化和2Gy照射后不同时间的p16基因转录水平的变化。结果 经研究证实,1～6Gy照射后p16的转录水平高于对照,在2～4Gy照射后达到峰值,2Gy照射后不同时间均有所增高,在照射后24h增高最明显。结论 2Gy60Coγ射线照射后p16基因转录水平在2～24h呈现时间依赖性的增高,在24h增高最明显,48和72h逐渐降低,但仍高于对照组。提示60Coγ射线可激活早期反应生长因子Egr-1,从而介导其下游基因p16的表达增强。 |
| 英文摘要: |
| Objective To construct a recombinant plasmid p-Egr- p16, investigate the expression of p16 induced by radiation in SMMC-7721 cells and its role in hepatoma suppression. Methods p-Egr-p16 vector that included Egr-1 and p 16 gene was constructed by restriction enzyme digest, ligation and introduced into SMMC-7721 cells by lipofectamine-mediated transfection. Semi-quantitati ve RT -PCR was used to detect the dose-effect and time-course manners of p16 mRNA l evel after irradiation. Results The results showed that p16 mRNA level was higher tha n that of the controls after 1-6.0 Gy irradiation. There was a peak after 2-4 Gy irradiation. p16 mRNA level increased at different times after 2 Gy irradiat ion, and the increase was marked at the 24th hour. Conclusions It is shown that p16 expression levels in SMMC-7721 c ells at 2-48 h after 2 Gy irradiation are higher than those of the sham-irradi at ion group. p16 expression levels show a time-dependant increase from 2 to 24 ho urs, and then a decrease at 48 and 72 h, but it is still higher than the sham-i rradiation group. It is suggested that 60Coγ-rays could activate Eg r-1 gene promoter and induce the expression of downstream gene p16. |
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