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Ha-ras基因转化的人乳腺上皮细胞的扫描显微光度图像分析

Scanning microphotometry image analysis of Ha-ras-transformed human breast epithelial cells.

作者信息

Mello M L, Lin T Y, Russo J

机构信息

Department of Cell Biology, Institute of Biology, UNICAMP/IB, Campinas, SP, Brazil.

出版信息

Anal Cell Pathol. 1994 Dec;7(4):301-19.

PMID:7696155
Abstract

Nuclei of the human breast epithelial cells, MCF-10A, transfected with the c-Ha-ras oncogene, the ras proto-oncogene and the plasmid Homer 6 only, were studied by image analysis after Feulgen staining. This material had been previously used to demonstrate that the experimental insertion of the activated c-Ha-ras oncogene into the DNA of the MCF-10A cells induces their tumoural properties. The ras-transformed nuclei of the MCF-10A cells exhibited differences in chromatin supraorganization in comparison with the nuclei of human breast carcinoma MCF-7 cells, or c-Ha-ras-transformed NIH/3T3 cells and, to a much lesser extent, with those of other MCF-10A transfectants and the non-transfected MCF-10A cells. All MCF-10A transfectants exhibited unravelling of both condensed and non-condensed chromatin, which, however, was less drastic in the ras-transformed MCF-10A cells. It is hypothesized that simultaneous to a general chromatin loosening as a response to foreign transfected DNA, a reverse mechanism may be elicited by ras transformation in the chromatin of the MCF-10A cells. The result in terms of elicited chromatin condensation was not as strong as that promoted in ras-transformed NIH/3T3 cells. Considering that early steps of tumour progression in vitro have previously been assumed to be involved in the ras-transformed MCF-10A cells, the differences in chromatin supraorganization of the ras-transformed MCF-10A cells as compared with MCF-7 cells are probably due to their different tumoural stages plus the putative effect of the transfected DNA vector on the transfected MCF-10A cells.

摘要

对转染了c-Ha-ras癌基因、ras原癌基因以及仅转染了质粒Homer 6的人乳腺上皮细胞MCF-10A的细胞核,在福尔根染色后进行图像分析研究。该材料先前已被用于证明将活化的c-Ha-ras癌基因实验性插入MCF-10A细胞的DNA中可诱导其肿瘤特性。与人类乳腺癌MCF-7细胞、c-Ha-ras转化的NIH/3T3细胞的细胞核相比,MCF-10A细胞的ras转化细胞核在染色质超组织方面表现出差异,与其他MCF-10A转染细胞和未转染的MCF-10A细胞的细胞核相比,差异程度要小得多。所有MCF-10A转染细胞均表现出凝聚和非凝聚染色质的解聚,然而,在ras转化的MCF-10A细胞中这种解聚程度较轻。据推测,作为对外源转染DNA的反应,在发生一般染色质松弛的同时,ras转化可能会在MCF-10A细胞的染色质中引发一种反向机制。在引发染色质凝聚方面的结果不如在ras转化的NIH/3T3细胞中那么强烈。鉴于先前认为体外肿瘤进展的早期步骤与ras转化的MCF-10A细胞有关,ras转化的MCF-10A细胞与MCF-7细胞在染色质超组织上的差异可能是由于它们处于不同的肿瘤阶段,再加上转染的DNA载体对转染的MCF-10A细胞的假定作用。

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