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波形蛋白和核纤层蛋白在人早幼粒细胞白血病细胞HL-60体外分化过程中的表达

Expression of vimentin and nuclear lamins during the in vitro differentiation of human promyelocytic leukemia cells HL-60.

作者信息

Paulin-Levasseur M, Giese G, Scherbarth A, Traub P

机构信息

Max-Planck-Institut für Zellbiologie, Ladenburg bei Heidelberg, Bundesrepublik Deutschland.

出版信息

Eur J Cell Biol. 1989 Dec;50(2):453-61.

PMID:2697560
Abstract

We have reported previously that the human promyelocytic leukemia cell line HL-60, in its undifferentiated state, is devoid of cytoplasmic intermediate filament proteins and nuclear lamins A and C, but does express lamin B. Using immunofluorescence and immunoblotting techniques, we have further investigated the expression of vimentin and lamins A and C during differentiation of these tumor cells along the macrophage or granulocytic pathway in response to the inducing effects of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) or dimethyl sulfoxide. Our results show that, while the expression of lamin B remains largely unchanged, the synthesis of vimentin and lamins A and C is dramatically enhanced during the maturation of HL-60 cells along both hemopoietic pathways. Northern blot analysis of cellular RNAs isolated from untreated and TPA-treated HL-60 cell populations as well as from control HeLa cells was performed using two oligonucleotides, one complementary to the 5' region common to human lamin A/C mRNAs and the other to the 5' region of hamster vimentin mRNA. Very low but still detectable amounts of vimentin and lamin A/C mRNAs were found in untreated HL-60 cell population, in accordance with the detection of small quantities of vimentin and lamins A and C in these populations. This is probably due to the presence of a small number of spontaneously differentiating cells. On the other hand, strong signals comparable to those obtained with RNA from control HeLa cells were detected for the three mRNA species from TPA-treated cells.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

我们之前报道过,人早幼粒细胞白血病细胞系HL-60在未分化状态下缺乏细胞质中间丝蛋白以及核纤层蛋白A和C,但确实表达核纤层蛋白B。利用免疫荧光和免疫印迹技术,我们进一步研究了在佛波酯12-O-十四酰佛波醇-13-乙酸酯(TPA)或二甲基亚砜的诱导作用下,这些肿瘤细胞沿巨噬细胞或粒细胞途径分化过程中波形蛋白以及核纤层蛋白A和C的表达情况。我们的结果表明,虽然核纤层蛋白B的表达基本保持不变,但在HL-60细胞沿两条造血途径成熟的过程中,波形蛋白以及核纤层蛋白A和C的合成显著增强。使用两种寡核苷酸对从未经处理和经TPA处理的HL-60细胞群体以及对照HeLa细胞中分离出的细胞RNA进行Northern印迹分析,其中一种寡核苷酸与人核纤层蛋白A/C mRNA共有的5'区域互补,另一种与仓鼠波形蛋白mRNA的5'区域互补。在未经处理的HL-60细胞群体中发现了非常低但仍可检测到的波形蛋白和核纤层蛋白A/C mRNA量,这与在这些群体中检测到少量波形蛋白以及核纤层蛋白A和C一致。这可能是由于存在少量自发分化的细胞。另一方面,对于经TPA处理的细胞的三种mRNA种类,检测到了与对照HeLa细胞RNA获得的信号相当的强信号。(摘要截短于250字)

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