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1.
Changes in proteoglycan and glycosaminoglycan (GAG) content and distribution may play an important role in the development of many diseases, atherosclerosis, cancer and diabetes. Human cell lines act as models for the underlying pathomechanisms. Despite the importance of proteoglycans for cell functioning, information on the GAG composition of most human cell lines is limited. Comparative analysis of the GAG Δdisaccharide amount in 22 human cell lines yielded a mean value of 94 ± 58 pmol/106 cells (mean ± SEM). Total GAG amount and heparan sulfate/heparin Δdisaccharide composition, but not chondroitin sulfate/dermatan sulfate Δdisaccharide composition, differed significantly between the investigated adherent and suspension cell lines. We provide a novel overview of GAG Δdisaccharide composition in 22 different human cell lines. 相似文献
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3.
New human breast cell lines were developed from metastatic breast cancer tissues and normal breast tissues. Primary cultures
were initiated from cellular outgrowths of explanted tissues or from mechanically isolated cells in two serum-free media.
Cell cultures derived from both cancer and normal tissues were immortalized with pRSV-T plasmid to generate permanent breast
cell lines that exhibited an epithelial morphology. Cell lines generated in this study were characterized with respect to
morphology, growth rate, karyotype, presence of specific genes, and the expression of epithelial and breast markers. The cell
lines expressed the epithelial cell markers, cytokeratins 8 and 18, and retained the capacity to produce human milk fat globulin.
They also express the BRCA-1, erbB2, and EGF receptor genes and possess the H-ras, K-ras, and p53 genes. Preliminary data
showed that one of the new cancer cell lines was highly sensitive to the cytotoxic action of taxol. It is envisioned that
the new breast cell lines will be useful as targets for identification of therapeutic agents against breast cancer and as
models for carcinogenesis studies.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
BackgroundNanotoxicology is a major field of study that reveals hazard effects of nanomaterials on the living cells.MethodsIn the present study, Copper/Copper oxide nanoparticles (Cu/CuO NPs) were prepared by the chemical reduction method and characterized by different techniques such as: X-Ray Diffraction, Transmission and Scanning Electron Microscopy. Evaluation of the toxicity of Cu/CuO NPs was performed on 2 types of cells: human lung normal cell lines (WI-38) and human lung carcinoma cell (A549). To assess the toxicity of the prepared Cu/CuOs NPs, the two cell types were exposed to Cu/CuO NPs for 72 h. The half-maximal inhibitory concentration IC50 of Cu/CuO NPs for both cell types was separately determined and used to examine the cell genotoxicity concurrently with the determination of some oxidative stress parameters: nitric oxide, glutathione reduced, hydrogen peroxide, malondialdehyde and superoxide dismutase.ResultsCu/CuO NPs suppressed proliferation and viability of normal and carcinoma lung cells. Treatment of both cell types with their IC50’s of Cu/CuO NPs resulted in DNA damage besides the generation of reactive oxygen species and consequently the generation of a state of oxidative stress.ConclusionOverall, it can be concluded that the IC50's of the prepared Cu/CuO NPs were cytotoxic and genotoxic to both normal and cancerous lung cells. 相似文献
5.
Summary Cell extracts of five mosquito cell lines and a tick cell line were examined for four cellular isozymes using a cellulose-acetate
electrophoretic technique. This method distinguished the cell lines that were derived from the different species. Intraspecies
distinctions were not made using the cell lines tested; the significance of this finding is discussed. The usefulness of this
technique in identifying a potentially mislabeled cell line was demonstrated.
This research was supported by contracts, DADA 17-72C-2170 of the U.S. Army and N00014-78C-0104 of the U.S. Office of Naval
Research and grants from the World Health Organization and the Rockefeller Foundation. 相似文献
6.
BACKGROUND: Glutathione plays crucial roles in antioxidant defence and glutathione deficiency contributes to oxidative stress and may therefore play a key role in the pathogenesis of many diseases. The objectives of the present study were to evaluate the effects on glutathione turnover of thiol and non-thiol antioxidants in human cell cultures and if any of the antioxidant had a short-term cellular effect against different levels of hydrogen peroxide. METHODS: We have investigated the effect on the total glutathione amount in HeLa and hepatoma cell cultures of thiol antioxidants in comparison with non-thiol antioxidants, such as a copper chelator, Vitamin C, and a flavonoid. Furthermore, we have investigated the short-term (within 24h) interaction of the different antioxidants with hydrogen peroxide. RESULTS AND CONCLUSION: Lipoic acid and quercetin (Quer) were the two antioxidants that showed the highest stimulation of glutathione synthesis in cell cultures as judged by the total glutathione amount. However, no antioxidant protected against hydrogen peroxide present in concentrations that lowered cell protein. This finding may be attributed to the fact that it is necessary to incubate cell cultures with antioxidants or small doses of oxidants for a period before the cultures are exposed to hydrogen peroxide in order to enhance the antioxidant defence. The presence of Quer and Vitamin C lowered cell protein and total glutathione even in cell cultures containing hydrogen peroxide in concentrations that did not lower cell protein. This finding might be attributed to pro-oxidant properties and formation of excess reactive oxygen species in the presence of Quer and Vitamin C. 相似文献
7.
Phenotypic characterization of telomerase-immortalized primary non-malignant and malignant tumor-derived human prostate epithelial cell lines 总被引:5,自引:0,他引:5
Gu Y Li H Miki J Kim KH Furusato B Sesterhenn IA Chu WS McLeod DG Srivastava S Ewing CM Isaacs WB Rhim JS 《Experimental cell research》2006,312(6):831-843
In vitro human prostate cell culture models are critical for clarifying the mechanism of prostate cancer progression and for testing preventive and therapeutic agents. Cell lines ideal for the study of human primary prostate tumors would be those derived from spontaneously immortalized tumor cells; unfortunately, explanted primary prostate cells survive only short-term in culture, and rarely immortalize spontaneously. Therefore, we recently have generated five immortal human prostate epithelial cell cultures derived from both the benign and malignant tissues of prostate cancer patients with telomerase, a gene that prevents cellular senescence. Examination of these cell lines for their morphologies and proliferative capacities, their abilities to grow in low serum, to respond to androgen stimulation, to grow above the agar layer, to form tumors in SCID mice, suggests that they may serve as valid, useful tools for the elucidation of early events in prostate tumorigenesis. Furthermore, the chromosome alterations observed in these immortalized cell lines expressing aspects of the malignant phenotypes imply that these cell lines accurately recapitulate the genetic composition of primary tumors. These novel in vitro models may offer unique models for the study of prostate carcinogenesis and also provide the means for testing both chemopreventive and chemotherapeutic agents. 相似文献
8.
Arndt NX Tiralongo J Madge PD von Itzstein M Day CJ 《Journal of cellular biochemistry》2011,112(9):2230-2240
Currently there is only a modest level knowledge of the glycosylation status of immortalised cell lines that are commonly used in cancer biology as well as their binding affinities to different glycan structures. Through use of glycan and lectin microarray technology, this study has endeavoured to define the different bindings of cell surface carbohydrate structures to glycan-binding lectins. The screening of breast cancer MDA-MB435 cells, cervical cancer HeLa cells and colon cancer Caco-2, HCT116 and HCT116-FM6 cells was conducted to determine their differential bindings to a variety of glycan and lectin structures printed on the array slides. An inverse relationship between the number of glycan structures recognised and the variety of cell surface glycosylation was observed. Of the cell lines tested, it was found that four bound to sialylated structures in initial screening. Secondary screening in the presence of a neuraminidase inhibitor (4-deoxy-4-guanidino-Neu5Ac2en) significantly reduced sialic acid binding. The array technology has proven to be useful in determining the glycosylation signatures of various cell-lines as well as their glycan binding preferences. The findings of this study provide the groundwork for further investigation into the numerous glycan-lectin interactions that are exhibited by immortalised cell lines. 相似文献
9.
Critical parameters influencing hyperthermia-induced apoptosis in human lymphoid cell lines 总被引:2,自引:0,他引:2
O'Neill KL Fairbairn DW Smith MJ Poe BS 《Apoptosis : an international journal on programmed cell death》1998,3(5):369-375
Brief mild hyperthermia is sufficient to induce apoptosis (programmed cell death) in many cell lines. Here we describe the effects of a number of factors modulating heat shock induced apoptosis outcomes. We report the effects of cell type, heat load, recovery times, cellular growth phase, and protein synthesis on the levels of apoptoses seen in heat stressed cell populations. We observe that a number of cell lines are competent to undergo heat stress induced apoptosis using both the comet assay and cellular and nuclear morphologies. Of the cell lines tested we saw a wide spectrum of sensitivities, ranging from resistant (less than 1% apoptotic after 12 h) to exquisitely sensitive (>95%). By incrementally increasing the heat load from 37–49°C, we observed a gradual increase in apoptosis with a significant change from apoptotic to necrotic death at temperatures beyond 45°C. The kinetics of the apoptotic response to heat shock were also examined. A time dependent increase in apoptotic cell death was seen after initial hyperthermic treatment with most cell types reaching a plateau at 18 h. In addition to these parameters we report that growth phase has a strong influence on the number of apoptoses induced as a result of heat stress. Cultured cells, grown to a plateau, undergo apoptosis at a much higher level than similarly treated cells taken during an exponential phase of growth. Finally, we determined the necessity of protein synthesis for apoptotic competency. 相似文献
10.
Philip S. Rudland Gillian E. Ollerhead Angela M. Platt-Higgins 《In vitro cellular & developmental biology. Animal》1991,27(2):103-112
Summary Transformation of primary cultures of human breast cells with simian virus 40 and clonal selection has yielded single-cell-cloned,
epithelial cell lines, as well as myoepithelial-related cell lines. When grown on floating collagen gels, the epithelial cell
lines give rise to branching rays of cells, thick fingerlike protrusions, saclike structures, and degenerating areas. The
myoepithelial-related cell lines give rise only to the branching rays. Epidermal growth factor stimulates the production of
the thick protrusions, whereas cholera toxin stimulates the production of the degenerating areas. Immunocytochemical staining
of these cultures using reagents directed against the cell surface-extracellular matrix or the cellular cytoskeleton confirms
the epithelial and myoepithelial nature of the cells, and demonstrates that the degenerating areas are undergoing squamous
metaplasia. The fingerlike protrusions consist of cords of cells composed of inner, epithelial and outer, myoepithelial-related
cells sometimes surrounding a central lumen reminiscent of ducts. The saclike structures resemble alveoli. Ultrastructural
analysis confirms the identification of the basic cell types and also identifies indeterminate cells possessing features of
both epithelial and myoepithelial cells. It is suggested that the epithelial cell lines represent human mammary stem cells
that can undergo processes of morphogenesis and differentiation in vitro to form many of the three-dimensional structures
found within the breast.
This work was supported by the North West Cancer Research Fund and the Cancer and Polio Research Fund. 相似文献
11.
Martin Müller Conrad Flössel Michael Haase Thomas Luther Sybille Albrecht Peter Paul Nawroth Youming Zhang 《Virchows Archiv. B, Cell pathology including molecular pathology》1993,64(1):265-269
Expression of tissue factor (TF), the cellular receptor of clotting factor VII/VIIa, is a feature of certain malignant tumours.
The TF gene has been classified as an immediate early gene responsive to serum and cytokines. Thus, the regulation of TF gene
expression seems to play a role in cell growth. Recently, we have shown that constitutive TF expression in MCF-7 breast cancer
cells is modulated by such growth factors as EGF, TGFα, and IL-1. The present study deals with the immunocytochemically detectable
cellular distribution of TF in human breast cancer cell lines MCF-7 and MaTu stimulated by EGF and TGFα. In MCF-7 cells growing
logarithmically, stimulation led to a significant increase of TF mRNA after 2 h (in situ hybridization, Northern blot) and
to maximum TF expression after 6 h (immunohistochemistry). When decorated by monoclonal antibodies, TF protein showed a pronounced
localization at ruffled membrane areas, cell edges, and processes of spreading cells after 6 and 20 h. In more flattened cells
TF was concentrated in peripheric lamellae and microspikes communicating with neighbouring cells. After epithelial colony
pattern had established, TF was predominantly accumulated at the intercellular boundaries. The vary same distribution patterns
as seen in MCF-7 cells were true for the stimulated MaTu cell line. The dynamics and cellular distribution patterns of stimulated
TF expression support the hypothesis that TF could be of importance for morphogenic events associated with the growth and
differentiation of breast cancer cells in culture. 相似文献
12.
Long-term human breast carcinoma cell lines of metastatic origin: Preliminary characterization 总被引:19,自引:0,他引:19
Cailleau Relda Olivé Matilde Cruciger Quita V. J. 《In vitro cellular & developmental biology. Plant》1978,14(11):911-915
Summary Nineteen human breast carcinoma cell lines have been established as continous cultures during the past 6 years in our laboratory.
This preliminary report is designed to list the lines by their designated code numbers (MDA-MB) and present a brief summary
of their morphological, cytogenetic and biochemical characteristics. Sixteen of our lines were obtained from pleural effusions,
two from brain metastases, and one from pericardial fluid. All lines have been shown to be distinct entities and are uncontaminated
by HeLa cells or each other. A lq marker chromosome is present in all but one of the lines examined.
This research was sponsored by the National Cancer Institute Contract NO1-CB-23869; Institutional Grant 5S 07 RR 5511-15 awarded
by the Division of Research Resources, and a Kelsey-Leary Grant NO 974. 相似文献
13.
Nayak SK Kakati S Harvey SR Malone CC Cornforth AN Dillman RO 《In vitro cellular & developmental biology. Animal》2000,36(3):188-193
Summary Cell lines are valuable resources for the study of the malignancy and potential therapy of human breast cancer. A major problem
with adapting fresh breast tumor specimens to grow in vitro is contamination by fibroblasts. Previously, we have reported
a technique to overcome this problem (Nayak, S. K; Dillman, R. O. Clin. Biotechnol. 3:237–242; 1991). We have recently established
two new breast cancer cell lines, HH315 and HH375, that were derived from abdominal and supraclavicular lymph node metastases
from two patients. They were characterized by (1) growth kinetics; (2) staining with monoclonal antibodies (MoAbs) to cytokeratin-19,
epithelial membrane antigen (EMA), anticarcinoembryonic antigen (CEA), breast cancer antigen 1 (BRST-1), breast cancer antigen
2 (BRST-2), Her2/neu, and p53; (3) expression of domains of urinary plasminogen activator (uPA), neural cell adhesion molecule
(NCAM), and haptoglobin (Hp) (Harvey et al., 1997); and (4) karyotypic analysis. Growth kinetic studies showed that doubling
times for both lines ranged from 48 to 96 h. These two cell lines were found to have characteristics of the metastatic breast
cancer cells. Both lines stained positive with MoAbs to cytokeratin-19 and EMA, thus confirming their epithelial origin. They
also strongly reacted with the pan-breast carcinoma MoAbs BRST-1 and BRST-2, and carcinoembryonic CEA MoAb. Both cell lines
overexpressed the oncogene proteins Her2/neu and p53. The tumor cells were negative for estrogen and progesterone receptors.
HH315 cells were poorly differentiated, whereas the HH375 cells exhibited adenocarcinoma morphology. Both cell lines showed
intense cell surface and some cytoplasmic staining for uPA, NCAM, and Hp domains, which is a characteristic of malignant neoplasms
(Harvey et al., 1997). The HH375 cell line showed two cell types, of which 60% were hyperdiploids with 60–70 chromosomes and
5–10 marker chromosomes. The remaining cells were polyploid with more than 200 chromosomes. Cell line HH315 consisted of only
a polyploid population. These cell lines may be useful in breast cancer research. 相似文献
14.
Fish cell lines: Establishment and characterization of nine cell lines from salmonids 总被引:3,自引:0,他引:3
C. N. Lannan J. R. Winton J. L. Fryer 《In vitro cellular & developmental biology. Plant》1984,20(9):671-676
Summary Nine permanent cell lines have been established from five species of salmonids native to America's Pacific Northwest. With
the exception of a hepatoma from an adult trout, the lines were derived from normal tissues of embryonic or juvenile fish.
Cells were routinely grown in Eagle's minimum essential medium with 10% fetal bovine serum. Optimum growth temperatures for
these lines ranged from 21 to 24°C. All survived storage for at least 1 yr at −65°C and at least 5 yr in liquid nitrogen.
Six of the lines were demonstrably free of any microbial contamination but mycoplasmas were found in three. Eight of the lines
were heteroploid. The morphology of only one was fibroblastic. All the lines effectively replicated one or more of the common
salmonid viruses. Isozyme patterns were consistent with those of the species of origin. These cell lines have significant
application in fish virology.
This work is a result of research sponsored in part by the Oregon State University Sea Grant College Program supported by
NOAA Office of Sea Grant, U.S. Department of Commerce, under Grant NA79AA-D-0016 and by the Oregon Department of Fish and
Wildlife under PL-89304 Anadromous Fish Act and is Oregon Agricultural Experiment Station Technical Paper 6857. 相似文献
15.
Vera Neumannova Des R. Richardson Karin Kriegerbeckova Jan Kovar 《In vitro cellular & developmental biology. Animal》1995,31(8):625-632
Summary Iron is essential for tumor cell growth. Previous studies have demonstrated that apart from transferrin-bound iron uptake,
mammalian cells also possess a transport system capable of efficiently obtaining iron from small molecular weight iron chelates
(Sturrock et al., 1990). In the present study, we have examined the ability of tumor cells to grow in the presence of low
molecular weight iron chelates of citrate. In chemically defined serum-free medium, most human tumor cell lines required either
transferrin (5 μg/ml) or a higher concentration of ferric citrate (500 μM) as an iron source. However, we have also found that from 13 human cell lines tested, 4 were capable of long-term growth
in transferrin-free medium with a substantially lower concentration of ferric citrate (5 μM). When grown in medium containing transferrin, both regular and low-iron dependent cell lines use transferrin-bound iron.
Growth of both cell types in transferrin medium was inhibited to a certain degree by monoclonal antibody 42/6, which specifically
blocks the binding of transferrin to the transferrin receptor. On the contrary, growth of low-iron dependent cell lines in
transferrin-free, low-iron medium (5 μM ferric citrate) could not be inhibited by monoclonal antibody 42/6. Furthermore, no autocrine production of transferrin was
observed. Low-iron dependent cell lines still remain sensitive to iron depletion as the iron(III) chelator, desferrioxamine,
inhibited their growth. We conclude that low-iron dependent tumor cells in transferrin-free, low-iron medium may employ a
previously unknown mechanism for uptake of non-transferrin-bound iron that allows them to efficiently use low concentrations
of ferric citrate as an iron source. The results are discussed in the context of alternative iron uptake mechanisms to the
well-characterized receptor-mediated endocytosis process. 相似文献
16.
17.
Three human epithelial cell lines (CaCo-2, HEp-2 and HeLa) implicated as potential targets for three Fusarium toxins were tested for the extent of survival on exposure to increasing toxin concentration and incubation periods. Cytotoxicity assay using 3(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) was carried out with deoxynivalenol (DON), T-2 toxins and zearalenone (ZON) on CaCo-2, HEp-2 and HeLa cell lines. Of the three cell lines used, HeLa was the most sensitive, eliciting cell death after 2 days exposure at 100 ng ml–1with T-2 toxin. HeLa was the only cell line to exhibit cytotoxicity towards ZON showing cell death at 1000 ng ml–1after 2 days which increased to 4 days, showing substantial cell death at 200 ng ml–1. HEp-2 was sensitive to DON showing cell death after 2 days (100 ng ml–1) with complete cell death occurring at 200 ng ml–1 after 4 days of exposure. Substantial cytoxicity of T-2 towards HEp-2 occurred after 2 days at 1000 ng ml–1 and complete cell death occurred with 100 ng ml–1 at day 4. The CaCo-2 cell line was generally resistant to the mycotoxins tested between 100 and 1000 ng ml–1. This study shows that cytotoxicity of Fusarium toxins to epithelium cell lines is concentration- and time- dependant and results from ZON–HeLa interaction indicate possible cell type-mycotoxin specificity. 相似文献
18.
Einar K. Rofstad Anne Wahl Marit E. Hystad Jahn M. Nesland Trond Stokke 《Virchows Archiv. B, Cell pathology including molecular pathology》1991,60(1):189-197
Four new human melanoma cell lines were established in monolayer culture from xenograft lines originating from different patients.
Several distinct characteristics of the source xenograft lines were retained in the cell lines, e.g., number of chromosomes,
DNA-index, and cell ultrastructure. Cell volume was generally larger for the cell lines than for the corresponding xenograft
lines, but the differences among the lines were similar in vitro and in vivo. The cell lines showed significant differences
in growth pattern, i.e., cell motility and degree of intercellular contact. Cell cycle time (Tc) during exponential growth ranged from 15 to 21 h. The differences among the lines in Tc were mainly due to differences in the duration of S. Growth fraction was close to 100% and cell loss was negligible during
exponential growth. Plating efficiency was 90–100% in the presence of feeder cells. The four cell lines represent a valuable
supplement to the xenograft lines for future studies of the cell biology, pathophysiology, metastatic behavior, and treatment
sensitivity of malignant melanoma. 相似文献
19.
Rui Yan Mark Ottenbreit Bharati Hukku Michael Mally Sharong Chou Joseph Kaplan 《In vitro cellular & developmental biology. Animal》1996,32(10):656-662
Summary Methods for monitoring cell line identification and authentication include species-specific immunofluorescence, isoenzyme
phenotyping, chromosome analysis, and DNA fingerprinting. Most previous studies of DNA fingerprinting of cell lines have used
restriction fragment length polymorphism analysis. In this study, we examined the utility of an alternative and simpler method
of cell line DNA fingerprinting—polymerase chain reaction (PCR) amplification of fragment length polymorphisms. Fourteen human
cell lines previously found by other methods to be either related or disparate were subjected to DNA fingerprinting by PCR
amplification of selected fragment length polymorphism loci. Cell identification patterns by this method were concordant with
those obtained by isoenzyme phenotyping and restriction fragment length polymorphism-DNA fingerprinting, and were reproducible
within and between assays on different DNA extracts of the same cell line. High precision was achieved with electrophoretic
separation of amplified DNA products on high resolution agarose or polyacrylamide gels, and with fragment length polymorphism
(FLP) loci-specific “allelic ladders” to identify individual FLP alleles. Determination of the composite fingerprint of a
cell line at six appropriately chosen fragment length polymorphism loci should achieve a minimum discrimination power of 0.999.
The ability of PCR-based fragment length polymorphism DNA fingerprinting to precisely and accurately identify the alleles
of different human cell lines at multiple polymorphic fragment length polymorphism loci demonstrates the feasibility of developing
a cell line DNA fingerprint reference database as a powerful additional tool for future cell line identification and authentication. 相似文献
20.
We investigated the molecular and pharmacologic characteristics of VIP receptors on two human SCLC cell lines: NCI-N592 and NCI-H345. With NCI-N592 cell, the order of potency of VIP-related peptides in inhibiting 125I-VIP binding and in stimulating cAMP production was typical of the human VIP receptor. By covalent cross-linking, a polypeptide of Mr 62,300 was obtained. Conversely, the behavior of NCI-H345 cell line was totally different: helodermin was the most potent peptide, VIP and PHI were equipotent, while hGRF and secretin were totally ineffective. These results suggest that NCI-N592 cells possess a typical VIP receptor while NCI-H345 cells possess a helodermin-preferring receptor, and that the natural target of helodermin might not be the VIP receptor. 相似文献