Tissue polypeptide antigen (TPA) is related to the non-epidermal keratins 8, 18 and 19 typical of simple and non-squamous epithelia: re-evaluation of a human tumor marker.

Weber, Klaus; Osborn, Mary; Moll, Roland; Wiklund, B.; Lüning, Björn

doi:10.1002/j.1460-2075.1984.tb02198.x

Cited by 137 publications

(50 citation statements)

References 38 publications

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“…The staining appears as a diffuse network for the entire cell population. A similar pattern was observed for both antigens which was expected due to the homology between TPA and cytokeratins (Weber et al, 1984) Figure 6c shows the immunolocalization of CEA in cell cultures of similar passage. All cells appeared positive for CEA, although not all of them show the same stain intensity.…”

Section: Immunocytochemistrysupporting

confidence: 81%

A new cell line (8701-BC) from primary ductal infiltrating carcinoma of human breast

Minafra

Morello²,

Glorioso³

et al. 1989

Br J Cancer

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Previous ultrastructural and biochemical studies on ductal infiltrating carcinomas (d.i.c.) of human breast have suggested that interactions between the tumour cells and extracellular collagenous matrix are important for the invasive behaviour of these neoplastic cells (Minafra et al., 1984a(Minafra et al., ,b, 1988Pucci Minafra et al., 1985. To examine this further we have attempted to establish a breast carcinoma cell line in vitro which retains many of the properties observed in vivo. To date the majority of breast cancerderived cell lines have been obtained from secondary tumours and pleural effusions (Dobrynin, 1963;Soule et al., 1973;Trempe & Fogh, 1973;Engel & Yung, 1978;Monaghan et al., 1985). Although a cell line derived from a primary breast carcinoma has recently been reported (Vandewalle et al., 1987) the relative paucity of cell lines derived from primary carcinomas (Hackett et al., 1977;Nordquist et al., 1975; Lasfargues et al., 1978;Rudland et al., 1985) may be related to the technical difficulties associated with the extraction of viable tumour cells from surrounding stroma. Here we report the isolation, establishment and characterisation of a continuous line of neoplastic cells isolated from a primary duct infiltrating carcinoma of human breast (8701-BC). These cells have been grown in monolayer cultures for more than 100 passages and have retained morphological characteristics similar to those of the original tumour. Materials and methods Tissue fractionationTumour fragments obtained from surgical operations and histologically diagnosed as ductal infiltrating carcinoma were washed under sterile conditions with a Ca2+ and Mg2+ -free balanced salt solution (BSS-CMF). Specimens to be used for cell culture were taken from the core of the tumour, cut into very small pieces and pre-incubated in BSS-CMF with gentle stirring at 37°C for 15 min. 8701-BC cell line was derived from a G2-G3 duct infiltrating carcinoma obtained from a 72-year-old patient, with extensive lymphonodal infiltration. Histological grading was evaluated according to Bloom & Richardson (1957).Cell culture Cells of primary cultures were grown in minimum essential medium (MEM) with Earle's salts (Biochrom) supplemented with 20% (v/v) FCS (Difco) and 10% (v/v) TPB (Difco). A cell density of 1-2 x 10s cells ml-1 was used to seed primary cultures in 5ml flasks. At confluency cells were subcultured following detachment by exposure to trypsin 0.5% (w/v) and Versene 0.04% (w/v)

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Section: Immunocytochemistrysupporting

confidence: 81%

A new cell line (8701-BC) from primary ductal infiltrating carcinoma of human breast

Minafra

Morello²,

Glorioso³

et al. 1989

Br J Cancer

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“…It was also noted that most breast carcinomas express phenotypic markers that are consistent with luminal origin. Furthermore, Weber et al [25] have colocalized the proposed tumor marker for breast cancer, tissue polypeptide antigen (TPA) with known cytoskeletal components such as cytokeratins 8, 18 and 19. Another study [26] has found specific staining for cytokeratin 7 and 20 in a study of 127 tumors.…”

Section: Resultsmentioning

confidence: 99%

An approach to the proteomic analysis of a breast cancer cell line (SKBR‐3)

Wu¹,

Hancock²,

Goodrich

et al. 2003

Proteomics

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An approach to the proteomic analysis of a breast cancer cell line (SKBR-3)This report describes the profiling of proteins in a sample prepared by laser capture microdissection (LCM) from a breast cancer cell line (SKBR-3). This experimental approach serves as a model system for proteomic studies on selected tissue samples and for studies of specific cell types. The captured cells were isolated in a dehydrated and reduced state and solubilized with a denaturing buffer. After dilution the protein mixture was digested with trypsin and the resulting peptide mixture was fractionated by reversed phase HPLC (RPLC) and analyzed on an ion trap mass spectrometer. A key part of this study is the combination of the LCM process with an extraction/digestion procedure that allowed effective solubilization of a significant part of the cellular sample in a single step. The identity of the peptides was determined by tandem mass spectrometry measurements in which the resulting spectra were compared with genomic and proteomic databases and protein identifications were made. While only peptides with a high probability assignment were used, the interpretation of mass spectral fragmentation patterns were also confirmed by manual interpretation of the spectra. Also, for the more abundant proteins the initial protein assignment from the best match peptide was strengthened by the observation of additional confirmatory peptide identifications. Another selection criteria was correlation of the mass spectrometric studies with clinical and genomic studies of potential cancer markers in tumor samples. This proteomic study allowed identification of the following proteins: human receptor protein kinase HER-2 or ERBB-2 and related kinases HER-3 and HER-4, the gene products from breast cancer type I and II susceptibility genes and cytoskeletal components such as cytokeratins 8, 18 and 19. Other proteins include fibroblast growth factor receptor variants (FGFR-2&4) and T-lymphoma invasion and metastasis inducing protein 1 (TIAM1). In addition several nonreceptor protein kinases YES, FAK and JAK-1 and 3 were identified. Since the study was performed on a limited number of cells (approximately 10 000) it raises the possibility of such studies being performed on individual patient samples prepared by needle biopsy.

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“…Deletion analysis coupled with the known lamin caspase cleavage site (43,44) pointed to Asp-237 as a cleavage site that was verified by the inability of caspases to cleave K18 Asp-237 3 Glu (42). Interestingly, keratin fragments have been identified in sera of patients who harbor any one of a variety of epithelial tumors (45,46) and may have diagnostic potential (47).…”

Section: Discussionmentioning

confidence: 99%

Apoptosis Generates Stable Fragments of Human Type I Keratins

Ku¹,

Liao²,

Omary³

1997

Journal of Biological Chemistry

218

152

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Type I and II keratins help maintain the structural integrity of epithelial cells. Since apoptosis involves progressive cell breakdown, we examined its effect on human keratin polypeptides 8, 18, and 19 (K8, K18, K19) that are expressed in simple-type epithelia as noncovalent type I (K18, K19) and type II (K8) heteropolymers. Apoptosis induces rapid hyperphosphorylation of most known K8/18 phosphorylation sites and delayed formation of K18 and K19 stable fragments. In contrast, K8 is resistant to proteolysis and remains associated with the K18 fragments. Transfection of phosphorylation/glycosylation-mutant K8 and K18 does not alter fragment formation. The protein domains of the keratin fragments were determined using epitope-defined antibodies, and microsequencing indicated that K18 cleavage occurs at a conserved caspase-specific aspartic acid. The fragments are found preferentially within the detergentinsoluble pool and can be generated, in a phosphorylation-independent manner, by incubating keratins with caspase-3 or with detergent lysates of apoptotic cells but not with lysates of nonapoptotic cells. Our results indicate that type I keratins are targets of apoptosis-activated caspases, which is likely a general feature of keratins in most if not all epithelial cells undergoing apoptosis. Keratin hyperphosphorylation occurs early but does not render the keratins better substrates of the downstream caspases. Intermediate filament (IF)1 proteins encompass the nuclear lamins and a large family of tissue-specific cytoplasmic proteins that include keratins in epithelial cells, desmin in muscle, neurofilaments in neuronal cells, and vimentin in mesenchymal cells (reviewed in Refs. 1-3). Keratins are the largest IF protein subgroup and consist of more than 20 polypeptides (K1-K20) that are divided into relatively acidic type I (K9 -K20) and basic type II (K1-K8) keratins (4, 5). All epithelial cells typically express at least one type I and one type II keratin, as noncovalent obligate heteropolymers, in an epithelial cell-type specific manner. For example, simple-type epithelia preferentially express K8/18 with various levels of K19 and K20 (6 -10), keratinocytes express K1/10 and/or K5/14 depending on their differentiation state within the epidermal layer, and corneal epithelial cells express K3/12 (4). Although the full scope of keratin function is not known, one clear keratin function is to help maintain epithelial cell integrity particularly upon cell stress. This role is supported by several animal studies and a growing list of human epidermal, oral, and ocular diseases that result from keratin mutations (11-16).Keratins undergo several modifications that are likely involved in regulating their function (reviewed in Ref. 17), with phosphorylation being the most studied (reviewed in 18, 19). For K8/18, the known in vivo phosphorylation sites include Ser-52/Ser-33 of K18 (20) 2 and Ser-23/Ser-431/Ser-73 of K8 (22, 23). Of note, keratin phosphorylation is highly dynamic (24 -27) and is modulated during several phy...

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Tissue polypeptide antigen (TPA) is related to the non-epidermal keratins 8, 18 and 19 typical of simple and non-squamous epithelia: re-evaluation of a human tumor marker.

Cited by 137 publications

References 38 publications

A new cell line (8701-BC) from primary ductal infiltrating carcinoma of human breast

A new cell line (8701-BC) from primary ductal infiltrating carcinoma of human breast

An approach to the proteomic analysis of a breast cancer cell line (SKBR‐3)

Apoptosis Generates Stable Fragments of Human Type I Keratins

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