| 摘要: |
| 目的：探讨转录因子GATA4和甲胎蛋白（AFP）在小鼠胎肝和肝癌细胞中的表达及二者相关性。方法：获取小鼠孕10.5 d、孕13.5 d、孕15.5 d、孕17.5 d、出生后0.5 d的肝组织，半定量RT-PCR检测GATA4和AFP mRNA的表达，并以成鼠肝、正常肝细胞株和肝癌细胞株作为对照。将小鼠GATA4真核表达载体成功转染正常肝细胞并鉴定，将多柔比星作用肝癌细胞，然后均进行以下指标监测：半定量RT-PCR法检测GATA4、AFP和胎蛋白转录因子（FTF）mRNA的表达，免疫印迹法检测GATA4和AFP蛋白的表达，ELISA法检测细胞上清AFP的含量。结果：随着胎肝细胞分化的不断成熟，GATA4和AFP的表达逐渐下降，至出生后0.5 d已降至成肝水平。GATA4、FTF和AFP在正常肝细胞不表达，而在肝癌细胞高表达；正常肝细胞转染GATA4后，FTF和AFP表达显著增高，其细胞上清液AFP含量亦明显增高。多柔比星作用肝癌细胞后，6 h内GATA4的表达首先降低；随着时间延长（≥12 h），FTF和AFP表达逐渐降低，细胞上清液AFP含量亦逐渐降低，差别具有显著的统计学意义。结论：GATA4在胎肝组织和肝癌细胞中高表达并可能对AFP也有一定的上调作用。 |
| 关键词: GATA4转录因子 甲胎蛋白；胚胎；肝；肝肿瘤 |
| DOI：10.3724/SP.J.1008.2007.01309 |
| 投稿时间：2007-07-02 |
| 基金项目:国家自然科学基金(90208026). |
|
| Expression of GATA4 and alpha fetoprotein in fetal mouse liver and hepatocarcinoma cells and their relationship |
| REN Chuan-lu1, JI Jian1, SHEN Hong-shi1, LIU Hou-qi2 |
| (1. Department of Laboratory Medicine, No. 100 Hospital of PLA, Suzhou 215007, China; 2. Department of Histology and Embryology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China) |
| Abstract: |
| Objective：To explore the expression of GATA4 and alpha-fetoprotein (AFP) in fetal liver and hepatocarcinoma cells and their correlation. Methods: Semi-quantitive RT-PCR was used to detect the expression of GATA4 and AFP in liver tissues from mouse of E 10.5 d to P 0.5 d, normal liver cells, and hepatocarcinoma cells. Eukaryotic expression vector of mouse GATA4 was used to transfect normal liver cell line and the product was identified. Doxorubicine was used to treat hepatocarcinoma cells; then semi-quantitive RT-PCR was used to determine the mRNA of GATA4, fetoprotein transcription factor(FTF) and AFP; Western blot was used to examine the expression of GATA4 and AFP; and ELISA was used to determine AFP in supernatants. Results: The expression of GATA4 and AFP decreased with the maturation of fetal liver cells and reached adult level 0.5 d after birth. GATA4, FTF and AFP were not detected in normal liver cells but were overexpressed in hepatocarcinoma cells. The expression of FTF and AFP was increased in normal liver cells after transfection of GATA4, and the level of AFP was also increased in the supernatant. GATA4 was sharply decreased in hepatocarcinoma cells within 6 h after doxorubicine treatment; the expression of FTF and AFP was also decreased in supernatants 12 h after doxorubicine treatment (P<0.05). Conclusion: GATA4 is highly expressed in fetal liver and hepatocarcinoma cells,which may contribute to the elevation of AFP expression. |
| Key words: GATA4 transcription factors alpha-fetoproteins embryo liver liver neoplasms |
