共查询到20条相似文献,搜索用时 15 毫秒
1.
Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10 总被引:51,自引:0,他引:51
H D Soule T M Maloney S R Wolman W D Peterson R Brenz C M McGrath J Russo R J Pauley R F Jones S C Brooks 《Cancer research》1990,50(18):6075-6086
Two sublines of a breast epithelial cell culture, MCF-10, derived from human fibrocystic mammary tissue exhibit immortality after extended cultivation in low calcium concentrations (0.03-0.06 mM) and floating transfers in low calcium (MCF-10F), or by trypsin-Versene passages in the customary (normal) calcium levels, 1.05 mM (MCF-10A). Both sublines have been maintained as separate entities after 2.3 years (849 days) in vitro and at present have been in culture for longer than 4 years. MCF-10 has the characteristics of normal breast epithelium by the following criteria: (a) lack of tumorigenicity in nude mice; (b) three-dimensional growth in collagen; (c) growth in culture that is controlled by hormones and growth factors; (d) lack of anchorage-independent growth; and (e) dome formation in confluent cultures. Cytogenetic analysis prior to immortalization showed normal diploid cells; although later passages showed minimal rearrangement and near-diploidy, the immortal cells were not karyotypically normal. The emergence of an immortal culture in normal calcium media was not an inherent characteristic of the original tissue from which MCF-10 was derived since reactivated cryo-preserved cells from cultures grown for 0.3 and 1.2 years in low calcium were incapable of sustained growth in normal calcium. 相似文献
2.
Transforming growth factor beta-regulated gene expression in a mouse mammary gland epithelial cell line 总被引:4,自引:0,他引:4
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Xie L Law BK Aakre ME Edgerton M Shyr Y Bhowmick NA Moses HL 《Breast cancer research : BCR》2003,5(6):R187-R198
Background
Transforming growth factor beta (TGF-β) plays an essential role in a wide array of cellular processes. The most well studied TGF-β response in normal epithelial cells is growth inhibition. In some cell types, TGF-β induces an epithelial to mesenchymal transition (EMT). NMuMG is a nontransformed mouse mammary gland epithelial cell line that exhibits both a growth inhibitory response and an EMT response to TGF-β, rendering NMuMG cells a good model system for studying these TGF-β effects. 相似文献3.
Previous results have documented that Na2SeO3 has a biphasic effect on the growth of mammary cells in vitro. In the experiments reported herein, the effects of selenium on several parameters of cell proliferation in the YN-4 mouse mammary cell line were investigated. The biphasic effect of selenium on cell growth was confirmed; i.e., 5 X 10(-8) M selenium stimulated cell growth, whereas 5 X 10(-6) M delayed cell growth and 5 X 10(-5) M was cytotoxic. The inhibition of cell growth by 5 X 10(-6) M selenium was reversible when this dose was removed from the growth medium. The increased cell growth at 5 X 10(-8) M selenium was reflected by an increased cell number, increased uptake of [3H]thymidine into DNA, increased DNA labeling index, and an increased rate of DNA synthesis. The decreased cell growth at 5 X 10(-6) M selenium was reflected by a decrease in all of these parameters of cell growth kinetics. The differential effects of selenium were manifested by 48 hr after addition of selenium to the cell culture medium. The results indicate that one of the mechanisms of selenium-mediated inhibition of carcinogenesis may be due to an inhibition of cell proliferation of responsive cells. 相似文献
4.
J H Schiller G Bittner L F Meisner T D Oberley D Norback M Schwabe C R Faltynek N Raab-Traub 《Leukemia》1991,5(5):399-407
Hairy cell leukemia is a rare, B-cell malignancy uniquely sensitive to the antitumor effects of alpha and beta interferons (IFN). In order to further study the effects of IFN in this disease, we derived a cell line (HC1) from the peripheral blood mononuclear cells of a patient with hairy cell leukemia (HCL). Cells exhibited the typical morphological features of HCL, including the characteristic cytoplasmic projections by light, transmission, and scanning electron microscopy. HC1 cells were of B-cell lineage, as evidenced by immunophenotypic analysis. Although originally TRAP positive, HC1 cells lost this biochemical marker following 3 months in culture. Monoclonality of the cell line was confirmed by a clonal karyotypic abnormality characteristic of B-cell malignancies, and the presence of a single, distinctive fused terminal EBV fragment. The cells formed colonies in soft agar and were tumorigenic in irradiated nude mice. HC1 cells were sensitive to the antiproliferative effects of IFN-a and IFN-beta, but only moderately sensitive to the growth inhibitory effects of IFN-gamma. Incubating the cells in the presence of Type 1 IFN resulted in stabilization of cell numbers, without cellular proliferation or loss. Cell cycle analysis revealed that IFN-alpha resulted in a build-up of cells in the S phase of the cell cycle, suggesting a cytostatic effect of IFN on the growth of these cells. The HC1 cell line provides a model system which will be useful for in vitro studies of the biology and treatment of this disease. 相似文献
5.
Methionine and serine in combination enhanced the inhibitoryeffect of selenite on cell growth and DNA synthesis of the MODmammary epithelial cell line. These amino acids also increasedthe levels of a 58-kd selenoprotein which has been correlatedwith selenite's effects in previous studies. The use of theamino acids accelerated the onset of inhibition of DNA synthesisby selenite and increased the rate of actual selenoprotein synthesis.The mechanism of enhancement of selenite's effects was possiblydue to the amino acids increasing the levels of essential precursors(i.e. seryltRNAUGA, HSe) needed for selenoprotein synthesis. 相似文献
6.
Sun Yi; Oberley Larry W.; Oberley Terry D.; Elwell James H.; Sierra-Rivera Elaine 《Carcinogenesis》1993,14(7):1457-1463
Normal embryonal mouse liver cells in culture were shown toundergo spontaneous transformation during prolonged subculture.The spontaneously transformed cells lost their anchorage dependence,as measured by a soft agar assay, and gave rise to tumors innude mice. Accompanying this transformation, the antioxidantenzymes, copper-and zinc-containing superoxide dismutase (CuZnSOD),manganese superoxide dismutase (MnSOD), catalase (CAT) and glutathionereductase, decreased significantly in activity; the declinein enzymatic activity of CuZnSOD, MnSOD and CAT was due to adecline in the levels of immunoreactive protein. These spontaneouslytransformed high passage in vitro liver cells appeared similarin morphology, antioxidant enzyme activity and tumorigenicityto their counterparts transformed by N-methyl-N-nitro-N-nitrosoguanidineand Simian virus 40. These data provide experimental evidencethat changes in antioxidant enzymes are associated with spontaneousin vitro cellular transformation of mouse embryonal liver cells. 相似文献
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8.
E Y Lasfargues B Kramarsky N H Sarkar J C Lasfargues N Pillsbury D H Moore 《Cancer research》1970,30(4):1109-1117
9.
Kothari MS Ali S Buluwela L Livni N Shousha S Sinnett HD Vashisht R Thorpe P Van Noorden S Coombes RC Slade MJ 《British journal of cancer》2003,88(7):1071-1076
Currently, the therapy for breast cancer is determined by immunohistochemical staining of the primary tumour for oestrogen receptor alpha (ERalpha). However, a proportion of ERalpha-positive patients fail to respond to tamoxifen and a proportion of ERalpha-negative patients show response. Here, we describe a novel procedure for the purification of malignant breast epithelial cells in an attempt to identify these patients at an early stage. Using this procedure, we are able to purify malignant cells to >90% purity as determined by immunohistochemical staining, cytology and fluorescent in situ hybridisation (FISH). While the malignant cells can be maintained in culture they do not proliferate in contrast to purified breast epithelial cells from reduction mammoplasties. Moreover, ERalpha and progesterone receptor (PR) expression is maintained in malignant cells, whereas normal epithelial cells rapidly lose ERalpha and PR. Functional studies were performed on the separated malignant cells in terms of their response to oestradiol and tamoxifen. Four out of the seven ERalpha-positive tumours showed a significant reduction in cell numbers after tamoxifen treatment compared to oestradiol, ERalpha negative tumours failed to show a response. We conclude that (a) it is possible to purify and maintain breast cancer cells for a sufficient period to permit functional studies and (b) ERalpha is retained in culture facilitating the use of these cells in studies of the mechanism of endocrine response and resistance in vitro. 相似文献
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11.
Shen KC Miller F Tait L Santner SJ Pauley R Raz A Tainsky MA Brooks SC Wang YA 《Breast cancer research and treatment》2006,98(3):357-364
Summary We report the establishment of a breast epithelial cell model that undergoes growth arrest at different stages of the cell cycle depending upon the DNA damaging agents encountered. Primary breast epithelial cells from normal reductive mammoplasty were grown in low-calcium culture medium. Free-floating cells under this condition were separated and used for establishment of the MCF-15 breast epithelial cell line. We found that MCF-15 breast epithelial cells display a superb response to different phases of the cell cycle arrest in response to various DNA damaging agents. Immunohistological analysis indicates that MCF-15 cells express cytokeratin 19, CD44, CXCR4, SDF-1, SPARC and vimentin. Although less than 5% of the MCF-15 cells expressed Muc-1 in culture, increased Muc-1 expression was observed in luminal epithelial cells along the newly formed lumen in xenografts. Furthermore, a small population of MCF-15 cells expressed estrogen receptor-α (ERα) in xenografts while ERα expression was not detected in monolayer culture. Therefore, the MCF-15 breast epithelial cell line possesses characteristics of breast progenitor cells and provides a good cell culture model for studying the response to DNA damage and the etiology of aggressive basal-like breast cancers. 相似文献
12.
A spontaneously transformed human fetal lung cell line (HFLT) was derived from a human fetal lung diploid fibroblast cell line (2BS) by continuous culture. The biological characteristics of this cell line were studied and compared with those of 2BS cells. Accompanying the morphological alteration, the growth rate of the transformed cell was accelerated and the maximum cell density was increased. The anchorage-independent growth was shown by its ability to form colonies in soft agar. The collagen synthesis phenotype of 2BS cells was changed. In addition, the increased chromosome number and the nodule formation after heterotransplantation were pathognomonic of malignant transformation. 相似文献
13.
W L Kirkland N S Yang T Jorgensen C Longley P Furmanski 《Journal of the National Cancer Institute》1979,63(1):29-41
Normal and malignant human mammary epithelial cells were placed in culture. Normal cells were recovered from late-lactation milk and breast fluids, and malignant cells were isolated from primary breast tumors by collagenase digestion. The concentration of cells obtained from breast fluid samples was inversely proportional to the volume of fluid secreted. Most of these cells adhered rapidly to the substrate, did not replicate, displayed Fc receptor-dependent phagocytic activity, and were thus identified as macrophages. The remaining cells grew out into large islands comprised of one or two distinct morphologic types of mammary epithelial cells. Optimum growth of these cells was obtained in medium buffered to pH 6.8, and the epidermal growth factor markedly prolonged the exponential growth phase of the cells. Two morphologically distinct populations of epithelial cells were also observed in cultures established from individual breast tumors. Growth of the malignant cells was relatively unaffected by the pH of the culture medium, and the cells were unresponsive to exogenously added hormones. Overgrowth of malignant epithelial cells in primary cultures by stromal fibroblasts was retarded by replacement of standard growth medium with fresh medium containing a serum substitute; growth of the malignant epithelial cells was unaffected. A feeder layer of mitomycin C-treated human fibroblasts increased the plating efficiency of both normal and malignant cells in primary culture and also facilitated passage of these cells to secondary and tertiary cultures. 相似文献
14.
Basal lamina formation by normal and transformed mouse mammary epithelial cells duplicated in vitro 总被引:6,自引:0,他引:6
Cells from low-passage (LP) cultures of a mouse mammary epithelial line (NMuMG cells) form a basal lamina when they are cultured on a type I collagen gel substratum. A high-passage (HP) strain of this line maintained the morphologic, serologic, and karyologic properties of the LP cells. For the determination of whether transformation of the NMuMG cells might lead to defects in the basal lamina, cells from LP cultures were compared in vivo and in vitro with cells of HP cultures for tumorigenicity, growth characteristics, and ability to form a lamina. The LP NMuMG cells had a typical epithelial morphology and showed no cytologic evidence of cancer. They formed an ultrastructurally normal continuous basal lamina in vivo when they were injected into athymic nude mice. In contrast, the HP cells were pleomorphic and highly invasive when injected into nude mice where they showed frequent and large basal lamina defects. These cells also accumulated only traces of lamina-like materials when cultured on a collagen gel, indicating that neoplastic transformation had markedly reduced the ability of NMuMG cells to form a basal lamina both in vivo and in vitro. Because the collagen gel culture system duplicated the in vivo situation with regard to basal lamina integrity, the basis for this lack of in vitro basal lamina formation may be physiologically relevant for the mechanism of malignant invasion. 相似文献
15.
16.
J Costa P M Howley F Legallais C Yee N Young A S Rabson 《Journal of the National Cancer Institute》1977,58(4):1147-1149
Primary cultures of NIH nude mouse (nu/nu) kidney cells were transformed with a human papovavirus (MMV). The transformed cell line expressed T-antigen, and MMV DNA was found to be associated with the cell DNA. When NIH nu/nu mice were inoculated with the transformed cells, they developed tumors at the injection site but failed to generate detectable levels of T-antibody. A control group of nu/+ littermates rejected the tumor inoculum but mounted an antibody response to T-antigen. It was proposed that nude mouse cells may be a suitable system to test oncogenicity of in vitro transformed cells. 相似文献
17.
The effects of selenium on three mammary epithelial cell lines (YN-4, WAZ-2t, and CL-S1) grown in vitro were examined by immunocytochemical and transmission electron microscopy technique. The primary effect of selenium at the ultrastructural level was the appearance of electron-dense inclusions within the mitochondrial matrix. The mitochondrial inclusions were seen in all three cell lines, although most readily induced in YN-4 cells, the cell line which is most sensitive to selenium-mediated growth inhibition. Selenium at 5 x 10(-8) and 5 x 10(-6) M did not alter cytoplasmic microtubules or intermediate filament networks, as determined by immunocytochemical staining. Immunocytochemical staining for cytoplasmic filaments and microtubules, and transmission electron microscopy observations, supported the contention that cells from all three cell lines were epithelial in origin, since they contained abundant desmosomes and were uniformly positive for keratin intermediate filaments. Whereas line YN-4 was negative for vimentin intermediate filaments, a minority (5 to 24%) of the cells in lines CL-S1 and WAZ-2t stained positively. In addition, the tumorigenicity of these three cell lines was assessed by in vitro growth assays and in vivo transplantation assays. Cell lines YN-4 and WAZ-2t, but not line CL-S1, were tumorigenic in syngeneic mice. All tumors were mammary adenocarcinomas. Cytochalasin B-induced multinucleation assay and growth as multicellular spheroides correlated positively with in vivo tumorigenicity, whereas saturation density and growth in low Ca2+ medium were not correlated with tumorigenicity. It is speculated that one of the early effects of selenium-mediated growth inhibition may be a modulation of mitochondrial function. 相似文献
18.
Esteban N Gurzov Sanaa M Nabha Hamilto Yamamoto Hong Meng O Graciela Scharovsky R Daniel Bonfil 《BMC cancer》2007,7(1):184
Background
Despite significant advancement in breast cancer therapy, there is a great need for a better understanding of the mechanisms involved in breast carcinogenesis and progression, as well as of the role of epigenetic contributions from stromal cells in mammary tumorigenesis. In this study, we isolated and characterized murine mammary tumor-derived epithelial and myofibroblast cell lines, and investigated the in vitro and in vivo effect of cellular soluble factors produced by the epithelial cell line on tumor cells. 相似文献19.
《European journal of cancer & clinical oncology》1982,18(4):399-403
Androgen responsive and unresponsive Shionogi 115 mouse mammary carcinoma cells have been examined for anchorage-independent growth and tumorigenicity in nude mice. The two cell types exhibit transformed and normal growth characteristics respectively, but both give rise to tumours in nude mice. No correlation between tumorigenicity and transformed characteristics including anchorage-in-dependent growth could be demonstrated. 相似文献
20.
Macrophage-mediated induction of drug-resistant variants in a mouse mammary tumor cell line 总被引:6,自引:0,他引:6
The ability of macrophages to induce drug-resistant variants was studied in an in vitro macrophage-tumor cell coculture system utilizing the hypoxanthine-guanine phosphoribosyl transferase locus as measured by resistance to 6-thioguanine. Tumor cells of mouse mammary tumor line 66 were sensitive to macrophage induction of thioguanine resistance as shown by an increase in the frequency of thioguanine-resistant variants which arose following macrophage coculture to levels at least 5- to 10-fold above the spontaneous frequency. Detection of increased numbers of variants depended upon the macrophage:tumor cell ratio, with 50:1 or greater being necessary. The activity of the macrophages was dependent upon their activation stage. The induction of drug-resistant variants could be inhibited by oxygen radical scavengers. The basis for the emergence of thioguanine-resistant cells appeared to be induction of new variants rather than selection of preexisting resistant cells from the parental population, since thioguanine-sensitive and -resistant cells were equally sensitive to macrophage-mediated toxicity. In six of the six macrophage-induced variants tested, resistance was associated with loss of hypoxanthine-guanine phosphoribosyl transferase activity. The reverse variation frequency at the hypoxanthine-guanine phosphoribosyl transferase locus in five macrophage-induced variants was low and similar to that of a stable ethyl methanesulfonate-induced, thioguanine-resistant line. Macrophages isolated directly from growing mammary tumors, as well as activated peritoneal macrophages, were capable of inducing thioguanine resistance in line 66 cells. 相似文献