Jacobs H, Birnie G D
Nucleic Acids Res. 1980 Jul 25;8(14):3087-103. doi: 10.1093/nar/8.14.3087.
Saturation hybridisation with labelled single-copy DNA shows that polysomal poly(A)+ RNA of rat liver and of a minimum-deviation rat hepatoma cell-line (HTC) have similar total complexities and that few sequences are specific to either cell-type. Hybridisation kinetics of polysomal cDNAs with template and heterologous cell RNAs indicate that a proportion of liver messengers are at greatly reduced abundance in the hepatoma, but not the converse. Hybridisations using fractionated cDNAs enriched for abundant polysomal sequences confirm these findings: on average, abundant liver mRNAs are about 100-fold rarer in hepatoma, whereas abundant hepatoma mRNAs are only 5-fold rarer in liver. This pattern is also implied by cell-free translation of polysomal poly(A)+ RNAs. The reactions of total and fractionated polysomal cDNA probes with poly(A)+ nuclear RNA indicate that there is much less disparity in abundance of these sequences at nuclear level, implying that the differences arise, at least in part, post-transcriptionally. We interpret the altered mRNA abundances in the hepatoma in terms both of its decreased functional specialisation and its ability to proliferate.
用标记的单拷贝DNA进行饱和杂交表明,大鼠肝脏和一种最小偏差大鼠肝癌细胞系(HTC)的多聚核糖体多聚腺苷酸(poly(A)+)RNA具有相似的总复杂度,并且很少有序列是这两种细胞类型所特有的。多聚核糖体cDNA与模板RNA和异源细胞RNA的杂交动力学表明,一部分肝脏信使RNA在肝癌中的丰度大大降低,但反之则不然。使用富集了丰富多聚核糖体序列的分级cDNA进行杂交证实了这些发现:平均而言,丰富的肝脏mRNA在肝癌中的稀有程度约为100倍,而丰富的肝癌mRNA在肝脏中的稀有程度仅为5倍。这种模式也通过多聚核糖体多聚腺苷酸(poly(A)+)RNA的无细胞翻译得到暗示。总多聚核糖体cDNA探针和分级多聚核糖体cDNA探针与多聚腺苷酸(poly(A)+)核RNA的反应表明,这些序列在核水平上的丰度差异要小得多,这意味着差异至少部分是在转录后产生的。我们从肝癌功能特化降低及其增殖能力的角度来解释肝癌中mRNA丰度的改变。