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[乳腺细胞系HC11中依赖表皮生长因子的上皮细胞在表皮生长因子刺激期间表现出高度同步的细胞周期]

[EGF-dependent epithelial cells of the mammary cell line HC11 demonstrate a high degree of synchronized cell cycle during stimulation with epidermal growth factor].

作者信息

Avrov K O, Aksenov N D, Nikol'skiĭ N N, Kornilova E S

机构信息

Institute of Cytology, Russian Academy of Sciences, St. Petersburg.

出版信息

Tsitologiia. 1998;40(11):958-63.

Abstract

The most popular object for studying endocytosis of EGF-receptor complexes, human epidermoid carcinoma A431, was shown to answer to EGF in high concentration (100 ng/ml) by growth inhibition, being indifferent to lower (0.1-1 ng/ml) concentrations. At the same time, cells NIH 3T3, expressing human EGF receptor (HER14), and epithelial mammary cells HC11 increased 14C-thymidine incorporation into DNA after EGF addition. However, for HER14 cells stimulatory effect of EGF was twice weaker than that induced by serum, whereas the effect of EGF on 14C-thymidine incorporation in DNA of cells HC11 was approximately 5 times stronger compared to serum. Therefore, cells HC11 may be referred to as EGF-dependent. Cell cycle analysis by fluorimetry showed that more than 90% of serum-starved HER14 and HC11 were in G0/G1. Within 19-20 h after stimulation by EGF 70-90% of HC11 cells and only 30-40% of HER14 cells were in S-phase. EGF removing from culture medium earlier than 9-11 h after stimulation blocked entering of HC11 cells into S-phase, whereas such EGF-dependent period was not found for cells HER14. Thus, synchronization of progression through early stages of cell cycle, stimulated by EGF and the presence of well defined "early" (EGF-dependent) and "late" (EGF-independent) phases, make cells HC11 convenient object for studying physiological role of EGF receptor complexes endocytosis.

摘要

研究表皮生长因子(EGF)受体复合物内吞作用时,最常用的细胞是人表皮样癌A431细胞。研究发现,高浓度(100 ng/ml)的EGF可抑制该细胞生长,而低浓度(0.1 - 1 ng/ml)的EGF对其生长无影响。同时,表达人EGF受体(HER14)的NIH 3T3细胞和乳腺上皮细胞HC11在添加EGF后,其14C - 胸腺嘧啶核苷掺入DNA的量增加。然而,对于HER14细胞,EGF的刺激作用比血清诱导的作用弱两倍,而EGF对HC11细胞DNA中14C - 胸腺嘧啶核苷掺入的作用比血清强约5倍。因此,HC11细胞可被认为是EGF依赖性的。通过荧光法进行的细胞周期分析表明,超过90%血清饥饿的HER14细胞和HC11细胞处于G0/G1期。在EGF刺激后的19 - 20小时内,70 - 90%的HC11细胞进入S期,而只有30 - 40%的HER14细胞进入S期。在刺激后9 - 11小时之前从培养基中去除EGF会阻止HC11细胞进入S期,而HER14细胞未发现这种EGF依赖性时期。因此,由EGF刺激引起的细胞周期早期进程的同步化以及明确的“早期”(EGF依赖性)和“晚期”(EGF非依赖性)阶段的存在,使得HC11细胞成为研究EGF受体复合物内吞作用生理功能的便利对象。

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