A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium

Briand, Per; Petersen, Ole W.; Deurs, Bo van

doi:10.1007/bf02623578

Cited by 211 publications

(181 citation statements)

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“…Nonneoplastic S1 HMT-3522 HMECs (Briand et al, 1987), between passages 52 and 60, were plated at 2.3 ϫ 10 4 cells/cm 2 for propagation as monolayers (two-dimentsional [2D] culture) on plastic (Falcon; BD Biosciences Discovery Labware, Bedford, MA) in chemically defined H14 medium (Blaschke et al, 1994;Plachot and Lelièvre, 2004). To induce acinar differentiation in 3D culture, S1 cells were plated in the presence of 5% drip laminin-rich extracellular matrix (Matrigel; BD Biosciences Discovery Labware), as described previously, and kept for 10 d in H14 medium ; the 3D cell culture procedure is reviewed in Lelièvre and Bissell, 2005).…”

Section: Cell Culture and Induction Of Acinar Differentiationmentioning

confidence: 99%

“…Nonneoplastic HMT-3522 S1 HMECs (Briand et al, 1987) were cultured as monolayers (2D culture) to produce a high number of nondifferentiated cells, and nuclear matrix fractions were prepared according to classical protocols (Nickerson et al, 1997). In this method, the supernatant obtained after DNase I digestion and the pellet obtained after incubation with 2 M NaCl correspond to the DNase I-sensitive fraction and the nuclear matrix fraction, respectively.…”

Section: A Fraction Of Numa Is Present In the Chromatin Compartment Dmentioning

confidence: 99%

“…However, there is evidence that NuMA colocalizes with the chromatin-associated architectural protein HMG I/Y (Tabellini et al, 2001)-thought to play a role in chromatin structure and transcriptional regulation-and also interacts with the putative transcription factor GAS41 (Harborth et al, 2000). Because these data raise the possibility that NuMA may be associated with the chromatin compartment, we assessed the nuclear compartmentalization of NuMA in the HMT-3522 breast model by subcellular fractionation and in situ degradation of DNA.Nonneoplastic HMT-3522 S1 HMECs (Briand et al, 1987) were cultured as monolayers (2D culture) to produce a high number of nondifferentiated cells, and nuclear matrix fractions were prepared according to classical protocols (Nickerson et al, 1997). In this method, the supernatant obtained after DNase I digestion and the pellet obtained after incubation with 2 M NaCl correspond to the DNase I-sensitive fraction and the nuclear matrix fraction, respectively.…”

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NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

Abad

Lewis

Mian

et al. 2007

MBoC

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The coiled-coil protein NuMA is an important contributor to mitotic spindle formation and stabilization. A potential role for NuMA in nuclear organization or gene regulation is suggested by the observations that its pattern of nuclear distribution depends upon cell phenotype and that it interacts and/or colocalizes with transcription factors. To date, the precise contribution of NuMA to nuclear function remains unclear. Previously, we observed that antibody-induced alteration of NuMA distribution in growth-arrested and differentiated mammary epithelial structures (acini) in threedimensional culture triggers the loss of acinar differentiation. Here, we show that in mammary epithelial cells, NuMA is present in both the nuclear matrix and chromatin compartments. Expression of a portion of the C terminus of NuMA that shares sequence similarity with the chromatin regulator HPC2 is sufficient to inhibit acinar differentiation and results in the redistribution of NuMA, chromatin markers acetyl-H4 and H4K20m, and regions of deoxyribonuclease I-sensitive chromatin compared with control cells. Short-term alteration of NuMA distribution with anti-NuMA C-terminus antibodies in live acinar cells indicates that changes in NuMA and chromatin organization precede loss of acinar differentiation. These findings suggest that NuMA has a role in mammary epithelial differentiation by influencing the organization of chromatin. INTRODUCTIONThe nuclear mitotic apparatus protein (NuMA) was first described in 1980 (Lydersen and Pettijohn, 1980) and observed to concentrate at the spindle poles during mitosis. Subsequently NuMA, variously named SPN antigen, p240 antigen, centrophilin, 1F1/1H1 antigen, SP-H antigen, and W1 antigen (Compton et al., 1991(Compton et al., , 1992Kallajoki et al., 1991Kallajoki et al., , 1993Maekawa et al., 1991;Tousson et al., 1991;Yang et al., 1992;Maekawa and Kuriyama, 1993;Tang et al., 1993), was shown to be a critical player in the formation and stabilization of the mitotic spindle, notably by associating with the minus end of spindle microtubules and interacting with dynein, dynactin, and LGN (Compton and Cleveland, 1993;Gaglio et al., 1995;Merdes et al., 2000;Du et al., 2001;Gehmlich et al., 2004). In addition to its location at the poles of the mitotic spindle, NuMA has been reported in the nucleus of both cycling and growth-arrested cells, as shown by immunostaining of cells in culture and tissue biopsy sections (Lelièvre et al., 1998;Merdes and Cleveland 1998;Gribbon et al., 2002;Taimen et al., 2004). However, a role for NuMA in interphase remains to be determined. The hypothesis that NuMA might organize nuclear structure (Compton and Cleveland, 1994) is supported by the existence of a long central coiled-coil region in the protein and the prevalence of NuMA in the nucleus upon detergent extraction. Furthermore, NuMA binds DNA matrix attachment regions (MARs) in vitro (Ludérus et al., 1994), and the cleavage of NuMA precedes DNA degradation during apoptosis (Weaver et al., 1996). The likelihood of a link...

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Section: Cell Culture and Induction Of Acinar Differentiationmentioning

confidence: 99%

Section: A Fraction Of Numa Is Present In the Chromatin Compartment Dmentioning

confidence: 99%

mentioning

confidence: 99%

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NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

Abad

Lewis

Mian

et al. 2007

MBoC

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“…Cells were further harvested, centrifuged, and treated with hypotonic solution as previously described. 25 The cell pellet was fixed in methanol: glacial acetic acid (3:1) dropped on cold slides and stained for Giemsa bands. Chromosome counts were done on well-spread metaphases.…”

Section: Karyotypingmentioning

confidence: 99%

Epithelial to Mesenchymal Transition in Human Breast Cancer Can Provide a Nonmalignant Stroma

Petersen¹,

Nielsen²,

Gudjonsson³

et al. 2003

The American Journal of Pathology

263

196

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A breast carcinoma biopsy showed cytochemical evidence of epithelial mesenchymal transition and an ␣-smooth muscle actin-positive stromal reaction. To study the lineage, and the nature of the cells in the stromal reaction, we derived a novel cell line, HBFL-1, from the explanted biopsy. HBFL-1 cells are immortal and exhibit a shared non-random X-chromosome inactivation pattern with the epithelial tumor of origin. Yet they closely resemble normal, finite-life-span fibroblasts by morphology, lack of tumor formation in nude mice, marker expression profile, protein pattern using two-dimensional gel electrophoresis and the ability to undergo myofibroblast conversion. HBFL-1 interacts reciprocally with tumor cells in collagen gel to induce activation of MMP2, leading to tumor-like behavior of epithelial colonies. In vivo, HBFL-1 cells resembled normal-derived myofibroblasts and conferred a significant 3.5-to 7-fold increase in MCF-7 tumor size in nude mice. However, that they were indeed not normal fibroblasts was revealed by residual keratin expression and formation of epithelial microfoci in a reconstituted basement membrane and in nude mice. We conclude that breast cancer can generate its own nonmalignant stroma and that one function for this is that of a reciprocal interac- Epithelial mesenchymal transition (EMT) was originally described as a normal developmental process. 1,2 Later, it was adopted as an explanation for mesenchymal conversion in a number of cultured epithelial cells. 3,4 In cancer, EMT generally depicts a more aggressive behavior of the tumor cells. 5,6 In breast cancer, EMT has been estimated to occur in as much as 18% of tumors in vivo. [7][8][9] Under these conditions EMT is defined as the occurrence of a variable proportion of tumor cells that express mesenchymal markers such as vimentin, tenascin and stromelysin-3. 7,10 In its most elaborate form, EMT-derived cells of mixed epithelial-mesenchymal breast tumors may be difficult to distinguish from resident normal stromal cells. 11 These tumors which are also referred to as carcinosarcomas or metaplastic carcinomas in particular offer an excellent opportunity to study the nature and the consequence of this subset of EMT. 5,6 Such studies, however, have been hampered by lack of representative cell lines most likely due to low frequency of overtly metaplastic carcinomas but also to difficulties in culturing breast cancer cells in general. In the present study we succeeded in isolating a mesenchymal-like cell line from a metaplastic human breast carcinoma. Clonality analysis revealed that the cell line and the epithelial tumor cells of origin had a common ancestor. Even though the cells were immortal and severely aneuploid, and exhibited a rudimentary epithelial phenotype in terms of keratin expression and formation of microfoci in Matrigel and in vivo, they nevertheless behaved remarkably like normal resident fibroblasts. In particular they responded to transforming growth factor-␤ (TGF-␤) by having ␣-smooth muscle actin (␣-sm actin) induced and t...

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“…Huma 7, a normal human mammary epithelial cell line that had been immortalised with SV40 virus (Rudland et al, 1989), Huma 123, a benign human mammary epithelial cell line, and Huma 109, a benign human mammary elongated myoepithelial-like cell line, derived from a primary cell culture of human fibrocystic disease displaying prominent epithelial hyperplasia (Briand et al, 1987;Ke et al, 1993), and the malignant human mammary epithelial cell lines, MCF-7, T47D, and ZR-75-1, derived from pleural effusions of breast cancer patients (Soule et al, 1973;Engel and Young, 1978) and MDA-MB-231 (Cailleau et al, 1974) were propagated in a humidified atmosphere of 95% air, 5% CO 2 at 378C as described below. The Huma 7 cell line was grown in DMEM, 5% (v v 71 ) foetal calf serum (FCS), 50 ng ml 71 each of insulin and hydrocortisone.…”

Section: Human Breast Cell Linesmentioning

confidence: 99%

Identification of mRNAs differentially-expressed between benign and malignant breast tumour cells

et al. 2002

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Two suppression subtracted cDNA libraries have been constructed, one containing cDNAs to mRNAs present at a higher level in a benign human breast tumour-derived cell line relative to the malignant mammary cell line, MCF-7, and the other containing cDNAs present at a higher level in the MCF-7 cells relative to the benign cells. Randomly-picked cloned DNAs have been sequenced yielding 29 and 128 different cDNAs from the benign and malignant libraries, respectively. Using reverse Northern hybridisation, 76% and 83% of the cDNAs were differentially expressed by greater than two-fold, whilst 14% and 11% of cDNAs in the respective libraries were differentially expressed by more than 15-fold. Amongst these were oestrogenresponsive cDNAs and expressed sequence tags. One such oestrogen-responsive expressed sequence tag, M41, is transcribed from a gene located on chromosome 21q22.3, within an intron of a larger gene. The M41 gene contains oestrogen response elements, one of which is associated with alu repeats. M41 mRNA is expressed at a statistically significantly higher level in human breast cancer specimens than in normal human breast and benign lesions. In carcinomas, its up-regulation is associated with the development of the malignant cell.

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A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium

Cited by 211 publications

References 35 publications

NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

Epithelial to Mesenchymal Transition in Human Breast Cancer Can Provide a Nonmalignant Stroma

Identification of mRNAs differentially-expressed between benign and malignant breast tumour cells

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