Croizat B, Berthelot F, Felsani A, Gros F
Eur J Biochem. 1977 Apr 1;74(2):405-12. doi: 10.1111/j.1432-1033.1977.tb11405.x.
We have analysed the poly(A)-containing RNA from neuroblastoma cells at two different developmental states: either as round, immature neuroblasts grown in suspension, or as differentiated cells exhibiting the morphological properties of mature neurons, when attached to a culture dish. Suspension-grown and monolayer cells were pulse-labelled with tritiated uridine. The profile of cytoplasmic poly(A)-containing RNA from suspension cells is highly heterogeneous with peaks ranging from 16-30 S. The profile obtained from differentiated cells appears somewhat distinct from the previous one. This is evidenced by a relative decrease in the 26-S peak and a virtual disappearance of the 16-S component. In order to compare the 'steady-state' patterns of poly(A)-containing RNA in these two developmental stages, polysomal RNA was prepared from unlabelled cells. Following sucrose gradient sedimentation, each fraction was hybridized to [3H]poly(U). Examination of the two RNA hybridization profiles reveals striking similarities suggesting that 'steady-state' messenger populations include, on the average, the same subspecies. The 16-S fraction, which was not observed after the pulse-labelling of the monolayer culture, is detected here by hybridization to [3H]poly(U) when using polysomal poly(A)-containing RNA from monolayer cells as substrate. These results suggest that terminal differentiation of neuroblastoma cells is not accompanied by major alterations of the transcription program and is paralleled by a marked stabilization of the 16-S species.
我们分析了处于两种不同发育状态的神经母细胞瘤细胞中含聚腺苷酸(poly(A))的RNA:一种是悬浮培养的圆形未成熟神经母细胞,另一种是附着在培养皿上、表现出成熟神经元形态特征的分化细胞。悬浮培养的细胞和单层细胞用氚标记的尿苷进行脉冲标记。悬浮细胞中细胞质含poly(A)RNA的图谱高度异质,峰值范围为16 - 30S。从分化细胞获得的图谱与前一种图谱有些不同。这表现为26 - S峰相对下降以及16 - S成分几乎消失。为了比较这两个发育阶段含poly(A)RNA的“稳态”模式,从未标记的细胞中制备多聚核糖体RNA。经过蔗糖梯度沉降后,将每个组分与[³H]聚尿苷(poly(U))杂交。对这两种RNA杂交图谱的检查揭示了惊人的相似性,表明“稳态”信使群体平均包含相同的亚群。在单层培养物脉冲标记后未观察到的16 - S组分,当以单层细胞含poly(A)的多聚核糖体RNA为底物与[³H]poly(U)杂交时在此处被检测到。这些结果表明,神经母细胞瘤细胞的终末分化并不伴随着转录程序的重大改变,而是与16 - S物种的显著稳定并行。