EFFECT OF HYPERTHERMIA ON THE PROTEIN-CONTENT OF HELA-CELL NUCLEI - FLOW CYTOMETRIC ANALYSIS
EFFECT OF HYPERTHERMIA ON THE PROTEIN-CONTENT OF HELA-CELL NUCLEI - FLOW CYTOMETRIC ANALYSIS
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DOI：
10.2307/3574973
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发表时间：
1979-01-01
影响因子：
3.4
通讯作者：
ROTIROTI, JL
中科院分区：
文献类型：
--
作者：
BLAIR, OC;WINWARD, RT;ROTIROTI, JL
先前的研究表明，与未加热的细胞相比，从加热的细胞中分离的染色质的蛋白质含量相对增加。使用流式细胞术和生化方法，观察到从加热的细胞中分离出的 HeLa [人宫颈癌]细胞核的蛋白质含量增加。 HeLa 细胞在 45°C 下加热 7.5、15 或 30 分钟。 C，细胞核分离、固定、用异硫氰酸荧光素 (FITC) 染色并用流式细胞仪进行分析。 45°C 加热 7.5、15 或 30 分钟的细胞的 FITC 染色细胞核的平均荧光通道。相对于对照的C为1.46±。 0.05, 1.88.+-. 0.14 和 2.02.+-。分别为0.14。标签研究表明 FITC 可能对赖氨酸不具有特异性。相对于对照核的平均光散射通道（角度为1-15°）为1.18±。 0.04, 1.34.+-. 0.08 和 1.46.+-。 0.10（分别在 45°C 下 7.5、15 或 30 分钟）。电子体积测定与显微镜测量体积相结合表明加热电池的原子核中的电阻降低。生化测定的加热细胞核蛋白含量的相对增加为1.18.+-。 0.04, 1.30.+-. 0.06 和 1.39.+-。 0.06 和 1.39.+-。 0.05（分别在 45°C 下 7.5、15 或 30 分钟）。假设非染色质核蛋白的贡献是恒定的，则热诱导的核蛋白含量增加可以通过在染色质中观察到的蛋白质含量增加来解释。
Previous studies have shown a relative increase in the protein content of chromatin isolated from heated cells compared with unheated cells. Flow cytometric and biochemical methods were used and an increased protein content of HeLa [human cervical carcinoma] nuclei isolated from heated cells was observed. HeLa cells were heated for 7.5, 15 or 30 min at 45.degree. C, nuclei isolated, fixed, stained with fluorescein isothiocyanate (FITC) and analyzed with a flow cytometer. The mean fluorescence channel of FITC-stained nuclei from cells heated for 7.5, 15 or 30 min at 45.degree. C relative to control was 1.46 .+-. 0.05, 1.88 .+-. 0.14 and 2.02 .+-. 0.14, respectively. Labeling studies suggest that FITC may not be specific for lysine. The mean light scatter channel (for angles of 1-15.degree.) relative to control nuclei was 1.18 .+-. 0.04, 1.34 .+-. 0.08 and 1.46 .+-. 0.10 (for 7.5, 15, or 30 min at 45.degree. C, respectively). Electronic volume determinations in conjunction with microscopically measured volume indicate that the electrical resistance is reduced in nuclei from heated cells. The relative increase in the nuclear protein content of heated cells determined biochemically was 1.18 .+-. 0.04, 1.30 .+-. 0.06 and 1.39 .+-. 0.06 and 1.39 .+-. 0.05 (for 7.5, 15, or 30 min at 45.degree. C, respectively). The heat-induced increase in nuclear protein content can be accounted for by the increased protein content observed in chromatin assuming the contribution from non-chromatin nuclear protein is constant.