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核仁素与蛋白质B23在循环癌细胞中的比例恒定定量传递。

Proportionally constant quantitative transmission of nucleolin and protein B23 in cycling cancer cells.

作者信息

Sirri V, Pession A, Trerè D, Montanaro L, Derenzini M

机构信息

Centro di Patologia Cellulare, Dipartimento di Patologia Sperimentale, Via San Giacomo 14, Bologna, Italy.

出版信息

Clin Mol Pathol. 1995 Oct;48(5):M264-8. doi: 10.1136/mp.48.5.m264.

Abstract

Objective-To investigate whether and to what extent the two major AgNOR proteins, nucleolin and protein B23, are maintained after one cell division in proliferating cells.Design-Using three asynchronously growing human cancer cell lines, TG, SJNKP, and CHP 212 cells, nucleolin and protein B23 were first identified on SDS-polyacrylamide separated nucleolar proteins, transferred to nitrocellulose and silver stained for AgNOR proteins. Measurement of doubling time indicated a period very close to 24h for each of the cell lines. To quantify the percentage of nucleolin and protein B23 maintained in daughter cells after duplication, cells were labelled with [(35)S]-methionine and a 24h cold chase performed. Nucleolin and protein B23 labelling was evaluated by densitometric analysis on nitrocellulose autoradiograms.Results-The radioactivity relative to nucleolin and protein B23 bands maintained in the daughter cells was a constant fraction of that present before cell duplication. In the three cell lines the percentage of residual radioactivity measured in the nucleolin bands was 42.2, 40.6, and 41.2 and in protein B23 bands 48.0, 46.2, and 44.1.Conclusions-After one cell division the nucleolin and protein B23 quantity present in cells may be highly variable, depending on the amount of the two proteins present in the mother cell. This is important in relation to the correct utilisation of AgNOR protein quantity as an index for evaluating cell kinetics.

摘要

目的——研究在增殖细胞进行一次细胞分裂后,两种主要的核仁组成区嗜银蛋白(AgNOR),即核仁素和蛋白质B23,是否以及在何种程度上得以保留。

设计——使用三种异步生长的人类癌细胞系,即TG、SJNKP和CHP 212细胞,首先在十二烷基硫酸钠-聚丙烯酰胺(SDS-聚丙烯酰胺)分离的核仁蛋白上鉴定核仁素和蛋白质B23,转移至硝酸纤维素膜上并进行银染以检测AgNOR蛋白。倍增时间的测量表明,每种细胞系的倍增时间非常接近24小时。为了量化复制后子细胞中保留的核仁素和蛋白质B23的百分比,用[³⁵S]-甲硫氨酸标记细胞并进行24小时冷追踪。通过对硝酸纤维素膜放射自显影片进行光密度分析来评估核仁素和蛋白质B23的标记情况。

结果——子细胞中保留的与核仁素和蛋白质B23条带相关的放射性是细胞复制前存在的放射性的恒定比例。在这三种细胞系中,核仁素条带中测得的残留放射性百分比分别为42.2%、40.6%和41.2%,蛋白质B23条带中分别为48.0%、46.2%和44.1%。

结论——在一次细胞分裂后,细胞中存在的核仁素和蛋白质B23的量可能高度可变,这取决于母细胞中这两种蛋白质的量。这对于正确利用AgNOR蛋白的量作为评估细胞动力学的指标具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8d9/407982/8950d1024059/clinmolpath00010-0049-a.jpg

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