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通过显微注射SV40 DNA永生化的人乳腺上皮细胞中11号染色体短臂(11p15)杂合性缺失。

Loss of heterozygosity for the short arm of chromosome 11 (11p15) in human milk epithelial cells immortalized by microinjection of SV40 DNA.

作者信息

Garcia I, Brandt D, Weintraub J, Zhou W G, Aapro M

机构信息

Division of Onco-hematology, University Hospital, Faculty of Medicine, Geneva, Switzerland.

出版信息

Cancer Res. 1991 Jan 1;51(1):294-300.

PMID:1703035
Abstract

We have developed, by microinjection of SV40 DNA into human milk epithelial cells, a new mammary cell line, Hu-MI, which exhibits the phenotype of luminal cells or so-called "breast cancer precursor cells." This cell line retains the phenotype of primary cells as demonstrated by the expression of keratins 18 and 19 and of polymorphic epithelial mucins. However, the cells do not grow in agar after more than 80 passages, nor do they form tumors in nude mice. Established cells contain 2 copies of SV40 DNA integrated into the cellular genome and up to 14 copies of free SV40 DNA. A deletion of the short arm of chromosome 11 (11p15) including the c-Ha-ras and the beta-globin genes was found in the immortalized cells when the DNA from these cells was compared to the DNA from peripheral blood mononuclear cells obtained from the same donor. In addition, this cell line showed a good transfection efficiency for other DNA sequences using classical transfection and selection techniques with a neomycin resistance gene (pKOneo). Selective microinjection of DNA into tumor precursor cells may prove useful for the study of the molecular mechanisms involved in breast carcinogenesis. The possible significance of the loss of 11p13-15 in malignant progression of breast cancer is discussed.

摘要

通过将SV40 DNA显微注射到人乳腺上皮细胞中,我们建立了一种新的乳腺细胞系Hu-MI,它表现出腔面细胞或所谓“乳腺癌前体细胞”的表型。该细胞系保留了原代细胞的表型,这通过角蛋白18和19以及多态性上皮粘蛋白的表达得以证明。然而,经过80多次传代后,这些细胞在琼脂中不再生长,也不会在裸鼠中形成肿瘤。已建立的细胞含有整合到细胞基因组中的2份SV40 DNA以及多达14份游离的SV40 DNA。当将这些细胞的DNA与从同一供体获得的外周血单个核细胞的DNA进行比较时,在永生化细胞中发现了包括c-Ha-ras和β-珠蛋白基因在内的11号染色体短臂(11p15)缺失。此外,使用带有新霉素抗性基因(pKOneo)的经典转染和筛选技术时,该细胞系对其他DNA序列显示出良好的转染效率。将DNA选择性显微注射到肿瘤前体细胞中可能对研究乳腺癌发生过程中涉及的分子机制有用。文中还讨论了11p13 - 15缺失在乳腺癌恶性进展中的可能意义。

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