(Cell line: HMEC)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): LTQ Orbitrap XL
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: Gabriela Chiosis
PROVIDER: MSV000079862 | MassIVE | Thu Jun 30 09:44:00 BST 2016
SECONDARY ACCESSION(S): PXD004479
REPOSITORIES: MassIVE
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Project description:Glioblastoma multiforme is a highly aggressive malignant primary brain tumor in humans, with poor prognosis. ANGM5 has been established from a cerebral glioblastoma multiforme in a 72-years-old Iraqi man who underwent surgery for an intracranial tumor in 2005. The morphology, growth kinetics, karyotype, immunocytochemistry and angiogenesis factors expression profile were studied. ANGM5 has been grown continuously for more than 200 serial passages in culture for the last 13 years. The cultured cells are elongated or multipolar in shape and the population doubling time is 28 hours. The karyotype is complex, and the chromosomal number varied between 39-114. ANGM5 is resistant to Temozolomide, cisplatin, vincristine and etoposide. This chemoresistance could be explained by the overexpression of the breast cancer resistance protein (BCRP) and MGMT. Immunocytochemistry analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP), and negative for nestin and calbindin. Protein microarray analysis showed high production of tissue inhibitor of metalloproteinase 2 (TIMP2) as well as other factors that are important for the invasiveness and aggressiveness of glioblastoma. ANGM5 is a useful addition to the cell lines currently available for study of the pathobiology and chemoresistance properties of glioblastoma multiforme and antitumor drug discovery. Angiogenesis signaling is being evaluated in new human Iraqi glioblastoma multiforme cell line. Here, we measured angiogenesis factors released by cancer cells in vitro. We find that glioblastoma up-regulates many proteins and metabolites during the logarithmic phase, suggesting initiation of their endo-vesiculo-membrane system. Importantly, GM-CSF and G-CSF as they increase angiogenesis through promoting endothelial cell function. Also, b-FGF which promote tumor growth and proliferation. Moreover, it showed up-regulation to inflammatory, and extracellular matrix proteins. These factors will help proliferating glioblastoma cells to overcome stress conditions, such as cytotoxic chemotherapy, serum deprivation, hypoxia in vitro and in vivo. These findings support other evidence of chemoresistance ability of this cell line named AMGN5 as it found to resist several types of conventional chemotherapies.
2018-08-24 | GSE118960 | GEO
Project description:The scientific community has responded to the misidentification of human cell lines with validated methods to authenticate these cells; however, few assays are available for nonhuman cell line identification. We have developed a multiplex polymerase chain reaction assay that targets nine tetranucleotide short tandem repeat (STR) markers in the mouse genome. Unique profiles were obtained from seventy-two mouse samples that were used to determine the allele distribution for each STR marker. Correlations between allele fragment length and repeat number were determined with DNA Sanger sequencing. Genotypes for L929 and NIH3T3 cell lines were shown to be stable with increasing passage numbers as there were no significant differences in fragment length with samples of low passage when compared to high passage samples. In order to detect cell line contaminants, primers for two human STR markers were incorporated into the multiplex assay to facilitate detection of human and African green monkey DNA. This multiplex assay is the first of its kind to provide a unique STR profile for each individual mouse sample and can be used to authenticate mouse cell lines.
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Project description:SummaryRecently, several high profile studies collected cell viability data from panels of cancer cell lines treated with many drugs applied at different concentrations. Such drug sensitivity data for cancer cell lines provide suggestive treatments for different types and subtypes of cancer. Visualization of these datasets can reveal patterns that may not be obvious by examining the data without such efforts. Here we introduce Drug/Cell-line Browser (DCB), an online interactive HTML5 data visualization tool for interacting with three of the recently published datasets of cancer cell lines/drug-viability studies. DCB uses clustering and canvas visualization of the drugs and the cell lines, as well as a bar graph that summarizes drug effectiveness for the tissue of origin or the cancer subtypes for single or multiple drugs. DCB can help in understanding drug response patterns and prioritizing drug/cancer cell line interactions by tissue of origin or cancer subtype.Availability and implementationDCB is an open source Web-based tool that is freely available at: http://www.maayanlab.net/LINCS/DCB CONTACT: avi.maayan@mssm.eduSupplementary informationSupplementary data are available at Bioinformatics online.
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Project description:Immortalized hepatic stellate cells (HSCs) established from mouse, rat, and humans are valuable in vitro models for the biomedical investigation of liver biology. These cell lines are homogenous, thereby providing consistent and reproducible results. They grow more robustly than primary HSCs and provide an unlimited supply of proteins or nucleic acids for biochemical studies. Moreover, they can overcome ethical concerns associated with the use of animal and human tissue and allow for fostering of the 3R principle of replacement, reduction, and refinement proposed in 1959 by William M. S. Russell and Rex L. Burch. Nevertheless, working with continuous cell lines also has some disadvantages. In particular, there are ample examples in which genetic drift and cell misidentification has led to invalid data. Therefore, many journals and granting agencies now recommend proper cell line authentication. We herein describe the genetic characterization of the rat HSC line HSC-T6, which was introduced as a new in vitro model for the study of retinoid metabolism. The consensus chromosome markers, outlined primarily through multicolor spectral karyotyping (SKY), demonstrate that apart from the large derivative chromosome 1 (RNO1), at least two additional chromosomes (RNO4 and RNO7) are found to be in three copies in all metaphases. Additionally, we have defined a short tandem repeat (STR) profile for HSC-T6, including 31 species-specific markers. The typical features of these cells have been further determined by electron microscopy, Western blotting, and Rhodamine-Phalloidin staining. Finally, we have analyzed the transcriptome of HSC-T6 cells by mRNA sequencing (mRNA-Seq) using next generation sequencing (NGS).
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Project description:A variety of analytical approaches have indicated that melanoma cell line UCLA-SO-M14 (M14) and breast carcinoma cell line MDA-MB-435 originate from a common donor. This indicates that at some point in the past, one of these cell lines became misidentified, meaning that it ceased to correspond to the reported donor and instead became falsely identified (through cross-contamination or other means) as a cell line from a different donor. Initial studies concluded that MDA-MB-435 was the misidentified cell line and M14 was the authentic cell line, although contradictory evidence has been published, resulting in further confusion. To address this question, we obtained early samples of the melanoma cell line (M14), a lymphoblastoid cell line from the same donor (ML14), and donor serum preserved at the originator's institution. M14 samples were cryopreserved in December 1975, before MDA-MB-435 cells were established in culture. Through a series of molecular characterizations, including short tandem repeat (STR) profiling and cytogenetic analysis, we demonstrated that later samples of M14 and MDA-MB-435 correspond to samples of M14 frozen in 1975, to the lymphoblastoid cell line ML14, and to the melanoma donor's STR profile, sex and blood type. This work demonstrates conclusively that M14 is the authentic cell line and MDA-MB-435 is misidentified. With clear provenance information and authentication testing of early samples, it is possible to resolve debates regarding the origins of problematic cell lines that are widely used in cancer research.
| S-EPMC5762610 | biostudies-literature
Project description:BackgroundIt is well established that cancer cells can fuse with endothelial cells to form hybrid cells spontaneously, which facilitates cancer cells traversing the endothelial barrier to form metastases. However, up to now, little is known about the biologic characteristics of hybrid cells. Therefore, we investigate the malignant biologic behaviors and proteins expression of the hybrid cell line EAhy926 with its parent cell line A549.MethodsCell counting and flow cytometry assay were carried out to assess cell proliferation. The number of cells attached to the extracellular matrix (Matrigel) was measured by MTT assay for the adhesion ability of cells. Transwell chambers were established for detecting the ability of cell migration and invasion. Tumor xenograft test was carried out to observe tumorigenesis of the cell lines. In addition, two-dimensional electrophoresis (2-DE) and mass spectrometry were utilized to identify differentially expressed proteins between in Eahy926 cells and in A549 cells.ResultsThe doubling time of EAhy926 cell and A549 cell proliferation was 25.32 h and 27.29 h, respectively (P > 0.1). Comparing the phase distribution of cell cycle of EAhy926 cells with that of A549 cells, the percentage of cells in G0/G1 phase, in S phase and in G2/M phase was (63.7% +/- 2.65%) VS (60.0% +/- 3.17%), (15.4% +/- 1.52%) VS (13.8% +/- 1.32%), and (20.9% +/- 3.40%) VS (26.3% +/- 3.17%), respectively (P > 0.05). For the ability of cell adhesion of EAhy926 cells and A549 cells, the value of OD in Eahy926 cells was significantly higher than that in A549 cells (0.3236 +/- 0.0514 VS 0.2434 +/- 0.0390, P < 0.004). We also found that the migration ability of Eahy926 cells was stronger than that of A549 cells (28.00 +/- 2.65 VS 18.00 +/- 1.00, P < 0.01), and that the invasion ability of Eahy926 cells was significantly weak than that of A549 cells (15.33 +/- 0.58 VS 26.67 +/- 2.52, P < 0.01). In the xenograft tumor model, expansive masses of classic tumor were found in the A549 cells group, while subcutaneous inflammatory focuses were found in the EAhy926 cells group. Besides, twenty-eight proteins were identified differentially expressed between in EAhy926 cells and in A549 cells by proteomics technologies.ConclusionAs for the biological behaviors, the ability of cell proliferation in Eahy926 cells was similar to that in A549 cells, but the ability in adhesion and migration of Eahy926 cells was higher. In addition, Eahy926 cells had weaker ability in invasion and could not form tumor mass. Furthermore, there were many differently expressed proteins between hybrid cell line Eahy926 cells and A549 cells, which might partly account for some of the differences between their biological behaviors at the molecular level. These results may help to understand the processes of tumor angiogenesis, invasion and metastasis, and to search for screening method for more targets for tumor therapy in future.
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Project description:EGFR-inhibition is required for targeted therapies of ERBB2-positive/EGFR high breast cancer. Approximately 30% of human ERBB2-positive breast tumors also express EGFR.
2014-08-13 | GSE50104 | GEO