Nahon J L, Gal A, Frain M, Sell S, Sala-Trepat J M
Nucleic Acids Res. 1982 Mar 25;10(6):1895-911. doi: 10.1093/nar/10.6.1895.
Rot analysis of hybridization data using highly labeled alpha-fetoprotein (AFP) and albumin (32P)cDNA probes has been used to quantitate AFP and albumin mRNA sequences in RNA preparations from different subcellular fractions of developing rat liver and Morris hepatoma 7777. In addition, size analysis of these mRNA sequences has been carried out by electrophoretic fractionation on agarose gels containing methylmercury hydroxyde and hybridization to radioactive cloned albumin and AFP cDNA probes. In all the tissues examined (fetal, newborn and adult rat liver, and hepatoma 7777) most of the albumin and AFP mRNA sequences were found associated with the polysomes as mature mRNA molecules; less than 2% of these sequences were present in the nuclear or the non polysomal cytoplasmic compartments. The number of AFP mRNA molecules was found to decrease in parallel in all the cellular compartments during rat liver development. In Morris hepatoma 7777 the content of albumin mRNA was considerably decreased in all the cellular fractions as compared to normal liver. These results demonstrate that post-transcriptional control mechanisms leading to an accumulation of non-functional mRNA molecules are not implicated in the changes of expression of albumin and AFP genes during rat liver development and neoplasia.
利用高标记的甲胎蛋白（AFP）和白蛋白（³²P）cDNA探针，对杂交数据进行Rot分析，已用于定量分析发育中大鼠肝脏和莫里斯肝癌7777不同亚细胞组分的RNA制剂中甲胎蛋白和白蛋白的mRNA序列。此外，这些mRNA序列的大小分析已通过在含有羟基汞甲基的琼脂糖凝胶上进行电泳分级分离，并与放射性克隆的白蛋白和AFP cDNA探针杂交来进行。在所检查的所有组织（胎儿、新生和成年大鼠肝脏以及肝癌7777）中，大多数白蛋白和AFP mRNA序列被发现以成熟mRNA分子的形式与多核糖体相关联；这些序列中不到2%存在于细胞核或非多核糖体细胞质区室中。发现在大鼠肝脏发育过程中，所有细胞区室中的AFP mRNA分子数量平行减少。与正常肝脏相比，在莫里斯肝癌7777中，所有细胞组分中的白蛋白mRNA含量均显著降低。这些结果表明，导致无功能mRNA分子积累的转录后控制机制与大鼠肝脏发育和肿瘤形成过程中甲胎蛋白和白蛋白基因表达的变化无关。
