Saha B K, Sameshima M, Sameshima F, Schlessinger D
In Vitro. 1981 Sep;17(9):816-24. doi: 10.1007/BF02618449.
Activities of three lysosomal enzymes--acid RNase. N-acetyl-beta-D-glucosaminidase and acid phosphatase--were determined during the growth cycles of WI-38 and HeLa cells, as well as in radiation-arrested WI-38 cells. In confluent and growth-arrested cultures of WI-38 cells, the lysosomal RNase increased six- to sevenfold; glucosaminidase, four- to fivefold; and phosphatase, two- to threefold. In HeLa cells, the lysosomal enzymes also increased in confluent cultures, but less than twofold; and the RNase level increased only transiently. In both WI-38 and HeLa cells, the rate of RNA breakdown also increased as cultures approached confluency. The rate of turnover of RNA, like the level of acid RNase, was higher in WI-38 cells than in HeLa cells (4 d half-life compared to 8 d). The increase in acid RNase could be prevented by incubation of cells in NH4Cl, but the rate of turnover in the presence of NH4Cl increased just as much when cells became confluent or stopped growth. The content of acid RNase could be changed more than 10-fold without altering the rate of RNA turnover. It is suggested that the increase in enzyme level is more important for possible autophagy or increased digestion of engulfed RNA, rather than for normal RNA turnover, when growth stops.
在WI-38和HeLa细胞的生长周期以及辐射阻滞的WI-38细胞中,测定了三种溶酶体酶——酸性核糖核酸酶、N-乙酰-β-D-氨基葡萄糖苷酶和酸性磷酸酶的活性。在汇合且生长停滞的WI-38细胞培养物中,溶酶体核糖核酸酶增加了6至7倍;氨基葡萄糖苷酶增加了4至5倍;磷酸酶增加了2至3倍。在HeLa细胞中,汇合培养物中的溶酶体酶也增加,但不到2倍;核糖核酸酶水平仅短暂增加。在WI-38和HeLa细胞中,随着培养物接近汇合,RNA分解速率也增加。RNA的周转速率,与酸性核糖核酸酶水平一样,WI-38细胞高于HeLa细胞(半衰期分别为4天和8天)。酸性核糖核酸酶的增加可通过将细胞在NH4Cl中孵育来阻止,但当细胞汇合或停止生长时,在NH4Cl存在下的周转速率增加幅度相同。酸性核糖核酸酶的含量可改变10倍以上而不改变RNA周转速率。这表明,当生长停止时,酶水平的增加对于可能的自噬或吞噬的RNA消化增加更为重要,而不是对于正常的RNA周转。