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Carcinoma Specimens (carcinoma + specimen)
Selected AbstractsQuantitative analysis of phosphopeptides in search of the disease biomarker from the hepatocellular carcinoma specimenPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 12 2009Hyoung-Joo Lee Abstract Reversible phosphorylation of proteins is the most common PTM in cell-signaling pathways. Despite this, high-throughput methods for the systematic detection, identification, and quantification of phosphorylated peptides have yet to be developed. In this paper, we describe the establishment of an efficient online titaniuim dioxide (TiO2)-based 3-D LC (strong cationic exchange/TiO2/C18)-MS3 -linear ion trap system, which provides fully automatic and highly efficient identification of phosphorylation sites in complex peptide mixtures. Using this system, low-abundance phosphopeptides were isolated from cell lines, plasma, and tissue of healthy and hepatocellular carcinoma (HCC) patients. Furthermore, the phosphorylation sites were identified and the differences in phosphorylation levels between healthy and HCC patient specimens were quantified by labeling the phosphopeptides with isotopic analogs of amino acids (stable isotope labeling with amino acids in cell culture for HepG2 cells) or water (HO for tissues and plasma). Two examples of potential HCC phospho-biomarkers including plectin-1(phopho-Ser-4253) and alpha-HS-glycoprotein (phospho-Ser 138 and 312) were identified by this analysis. Our results suggest that this comprehensive TiO2 -based online-3-D LC-MS3 -linear ion trap system with high-throughput potential will be useful for the global profiling and quantification of the phosphoproteome and the identification of disease biomarkers. [source] PEAZ-1: A new human prostate neoplastic epithelial cell lineTHE PROSTATE, Issue 2 2001Monika Schmelz Abstract BACKGROUND The generation of prostatic cell lines provides in vitro models for experimental studies of the pathogenesis of prostate carcinoma. Therefore, we established and characterized a new human prostate epithelial cell line, PEAZ-1 (prostate epithelial Arizona-1). METHODS The PEAZ-1 cells were grown from a primary human prostate carcinoma specimen obtained from radical prostatectomy. The isolated cells were characterized by immunobiochemistry, immunohistochemistry, and tumorigenicity studies. RESULTS PEAZ-1 cells are near diploid, tumorigenic, and androgen independent for cell growth. PEAZ-1 cells express N-cadherin, ,- and ,-catenins, and p120 at cell,cell contacts, cytoplasmic laminin 5, vinculin, paxillin, and phosphotyrosine at focal adhesions, vimentin, and cytokeratins 8 and 18. They do not express plakoglobin, E-cadherin, and PSA, and do not form desmosomes and hemidesomomes. PEAZ-1 respond to ocadaic acid, a pro-apoptotic agent, by expression of p53. CONCLUSIONS PEAZ-1 cells is a human prostate cancer cell line that has a number of mesenchymal characteristics. Prostate 48:79,92, 2001. © 2001 Wiley-Liss, Inc. [source] Expression and immunogenicity of NY-ESO-1 in hepatocellular carcinomaJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 8 2006Shinichiro Nakamura Abstract Background and Aim:, The present study was designed to investigate the expression of and humoral response against NY-ESO-1 in patients with hepatocellular carcinoma and to analyze the relationship between expression of NY-ESO-1 mRNA and clinicopathological features. Methods:, NY-ESO-1 mRNA and protein expression in surgically resected hepatocellular carcinoma specimens, adjacent non-cancerous liver and non-tumor bearing liver were examined by reverse transcription-polymerase chain reaction and immunohistochemical staining using a monoclonal antibody against NY-ESO-1 (ES121), respectively. The antibody response to NY-ESO-1 was examined by enzyme-linked immunosorbent assay using recombinant NY-ESO-1 protein. Results:,NY-ESO-1 mRNA was detected in 18 of 41 (43.9%) hepatocellular carcinomas. No NY-ESO-1 mRNA was expressed in 41 paired non-cancerous specimens and 18 specimens histologically diagnosed as liver cirrhosis or chronic hepatitis. Immunohistochemistry revealed heterogeneous expression of NY-ESO-1 protein in three of 18 NY-ESO-1 mRNA-positive hepatocellular carcinomas. None of 23 NY-ESO-1 mRNA-negative hepatocellular carcinomas expressed NY-ESO-1 protein. Antibody against NY-ESO-1 protein was detected in two of 92 patients with hepatocellular carcinoma. Both of these patients had tumors invading main branches of the portal vein. Conclusions:, The present study has demonstrated the expression of NY-ESO-1 mRNA in hepatocellular carcinoma and NY-ESO-1 antibody production in patients with advanced hepatocellular carcinoma. Although the enhancement of NY-ESO-1 protein expression and the activation of immune response of the patients with hepatocellular carcinoma are necessary, NY-ESO-1 has the potential to be a good target molecule for immunotherapy against advanced hepatocellular carcinoma. [source] AgNOR count as objective marker for dysplastic features in oral leukoplakiaJOURNAL OF ORAL PATHOLOGY & MEDICINE, Issue 9 2002Amit Chattopadhyay Abstract Background:,, Dysplasia is an important feature of leukoplakia. Because agreement among oral pathologists is poor regarding lesional diagnosis, silver stainable nucleolar organizer regions (AgNORs) as replicatory markers may have a place in objectively characterizing dysplasia in tissue specimens. Methods:,, We studied 41 normal oral epithelia, 51 oral leukoplakia (26 dysplastic, 25 non-dysplastic), and 51 cases of squamous cell carcinoma specimens for their mean AgNOR counts. Results:,, Mean AgNOR counts increased gradually from normal epithelium to non-dysplastic to dysplastic leukoplakia to squamous cell carcinoma. Using ROC analysis, we determined a mean AgNOR count cut-point (2.37) that can be used to distinguish between dysplastic and non-dysplastic leukoplakia. The test had a sensitivity of 75% and specificity of 83% with area under the curve being 88%. Conclusions:, Mean AgNOR count could be a valuable criterion for defining objective parameters for diagnosis/determination of dysplasia distinguishing between dysplastic and non-dysplastic leukoplakia. [source] Association between activation of atypical NF-,B1 p105 signaling pathway and nuclear ,-catenin accumulation in colorectal carcinomaMOLECULAR CARCINOGENESIS, Issue 2 2010Johannes C. Lauscher Abstract Recent studies have demonstrated that increased expression of coding region determinant-binding protein (CRD-BP) in response to ,-catenin signaling leads to the stabilization of ,-TrCP1, a substrate-specific component of SCF E3 ubiquitin ligase complex, resulting in an accelerated degradation of I,B, and activation of canonical nuclear factor-,B (NF-,B) pathway. Here, we show that the noncanonical NF-,B1 p105 pathway is constitutively activated in colorectal carcinoma specimens, being particularly associated with ,-catenin-mediated increased expression of CRD-BP and ,-TrCP1. In the carcinoma tissues exhibiting high levels of nuclear ,-catenin the phospho-p105 levels were increased and total p105 amounts were decreased in comparison to that of normal tissue indicating an activation of this NF-,B pathway. Knockdown of CRD-BP in colorectal cancer cell line SW620 resulted in significantly higher basal levels of both NF-,B inhibitory proteins, p105 and I,B,. Furthermore decreased NF-,B binding activity was observed in CRD-BP siRNA-transfected SW620 cells as compared with those transfected with control siRNA. Altogether, our findings suggest that activation of NF-,B1 p105 signaling in colorectal carcinoma might be attributed to ,-catenin-mediated induction of CRD-BP and ,-TrCP1. © 2009 Wiley-Liss, Inc. [source] Glycogen synthase kinase-3, does not correlate with the expression and activity of ,-catenin in gastric cancerAPMIS, Issue 10 2010YU-JIN CHO Cho Y-J, Yoon J, Ko Y-S, Kim S-Y, Cho S-J, Kim W-H, Park J-W, Youn H-D, Kim J-H, Lee B-L. Glycogen synthase kinase-3, does not correlate with the expression and activity of ,-catenin in gastric cancer. APMIS 2010; 118: 782,90. The regulation of ,-catenin activation by glycogen synthase kinase-3, (GSK-3,) in cancer has been shown to be cell type-specific. This study was performed to investigate the relationship between activated GSK-3, (phosphorylated at Tyr216) and ,-catenin in gastric cancer. Immunohistochemical tissue array analysis of 278 human gastric carcinoma specimens showed positive immunoreactivity for activated GSK-3, in 44% of the samples, whereas membranous ,-catenin and nuclear ,-catenin were observed in 19% and 20% of the samples, respectively. However, GSK-3, activation was not correlated with the expression of either membranous ,-catenin or nuclear ,-catenin. Moreover, SNU gastric cancer cell lines over-expressing kinase dead GSK-3, and the same cells treated with a GSK-3, inhibitor showed that GSK-3, inhibition did not alter either the protein expression or transcriptional activity of ,-catenin. In addition, GSK-3, activation was positively correlated with the expressions of anti-adenomatous polyposis coli (p = 0.002), p16 (p < 0.001), p21 (p < 0.001), p27 (p = 0.001), and p53 (p = 0.013). On the other hand, the nuclear expression of ,-catenin was positively correlated with those of Bcl-2 (p = 0.025) and cyclin D1 (p = 0.043), but these expressions were not correlated with GSK-3, activation. Thus, the GSK-3, pathway seems to function in gastric cancer cells without involving the ,-catenin pathway. [source] HER2/neu (c-erbB-2) gene amplification and protein expression are rare in uterine cervical neoplasia: a tissue microarray study of 814 archival specimensAPMIS, Issue 10 2009IANA LESNIKOVA Published studies have reported widely variable incidence of HER2/neu (c-erbB-2) protein expression and HER2/neu (c-erbB-2) gene amplification in cervical carcinoma. We examined tissue microarrays (TMAs) constructed from 814 formaldehyde-fixed paraffin-embedded archival specimens of cervical intraepithelial neoplasia (CIN)1 (n = 262), CIN2 (n = 230), CIN3 (n = 186) and invasive carcinoma (n = 136), for HER2/neu protein expression by immunohistochemistry (IHC) and for HER2/neu gene amplification by chromogenic in situ hybridization (CISH). We found moderate or strong immunohistochemical positivity for HER2/neu in 64 of 814 specimens (7.9%). Using CISH, polysomy of the HER2/neu gene was detected in 87 cases (10.7%), low/borderline amplification in five cases (0.6%) and true amplification in four cases (0.5%). The correlation between IHC and CISH was statistically significant in CIN2, CIN3 and invasive cervical carcinoma specimens. When present, Her-2/neu positivity is more commonly seen in higher grades of cervical dysplasia and in carcinoma. However, this large TMA study shows that HER2/neu oncoprotein expression and HER2/neu gene amplification overall are uncommon events in cervical neoplasia. This provides compelling evidence that HER2/neu plays no major role in the development and progression of cervical neoplasia. [source] Primary colorectal carcinomas and their intrapulmonary metastases: Clinical, glyco-, immuno- and lectin histochemical, nuclear and syntactic structure analysis with emphasis on correlation with period of occurrence of metastases and survivalAPMIS, Issue 6 2002Klaus Kayser Background. The aim of the study was to correlate clinical factors (disease-free interval/survival) with growth pattern in terms of structural entropy of patients with primary colorectal carcinomas and secondary lung lesions. Methods. Proliferation and apoptosis markers as well as determinants involved in information transfer by protein-carbohydrate interactions were monitored. The clinical history, surgical and histopathological reports, tumor load, survival of the patients with a maximum follow-up of 14 years, and sections of paraffin blocks of 60 colorectal carcinoma specimens and their pulmonary metastases were examined. Measurements of the staining intensities after processing sections of primary and secondary carcinomas with the marker panel and calculations of syntactic structure and stereological parameters were performed. Results. The majority of primary tumors (80%, 49/60) were surgically treated at advanced tumor stages (pT3/pT4), with detectable lymph node involvement (34/60). Lung metastases were resected after a median disease-free interval of 30.5 months, an average of 3.0 metastases adding up to a mean intrapulmonary tumor load of 9.98 ccm. The median survival was calculated to be 82 months after resection of the colon/rectal carcinomas and 40 months after that of intrapulmonary metastases. It was correlated with certain structural and vascular features such as vascular circumference. The proliferation index and several textural features were strongly associated with vascularization in primary and secondary tumors. Conclusions. Despite intra- and interindividual variations, vascularization properties and features such as bcl-2 positivity and CEA- and galectin-3-associated structural entropy in primary tumors or metastases are described as independent prognostic features. Absence of lymph node involvement or limited tumor stages of colon/rectal carcinomas should not exclude patients from thorough postsurgical scrutiny to detect lung metastases. [source] Angiotropism of human prostate cancer cells: implications for extravascular migratory metastasisBJU INTERNATIONAL, Issue 7 2005Claire Lugassy OBJECTIVES To report several samples of invasive human prostate cancer showing angiotropism, and to use human prostate cancer cells stably expressing green fluorescence protein (GFP) in in vitro and in vivo models to assess the dissemination pathway of prostate cancer cells. MATERIALS AND METHODS Malignant melanoma and prostate carcinoma cells can migrate along anatomical structures such as nerves; previous studies showed that melanoma cells can be perivascular, on the outside of the endothelium, i.e. they are angiotropic, which suggests the hypothesis that melanoma cells also may migrate along vascular channels, termed ,extravascular migratory metastasis' (EVMM). Thus we examined histologically 10 human prostatic carcinoma specimens for the presence of angiotropism. In vitro, the PC-3 prostate cancer cells were co-cultures with capillary-like structures. In vivo, PC-3 cells were implanted on the chick chorio-allantoic membrane (CAM). RESULTS Histologically, in all 10 cases, angiotropism was detected at least focally within the tumour or at the advancing front of the tumour. In vitro, the PC-3 cells spread along the external surface of the vascular tubules; in vivo, PC-3 cells formed a cuff around some vessels a few millimetres beyond the tumour, showing angiotropism. Histopathology of the CAM confirmed the perivascular location of tumour cells and the absence of tumour cells within the vessel lumina. CONCLUSION The presence of angiotropic tumour cells in human invasive prostate cancers, associated with the angiotropism of GFP prostate cancer cells cultivated in vitro and in vivo in angiogenic models, raises the possibility that some prostate tumour cells may migrate along the external surface of vessels as a mechanism of spread, i.e. EVMM. [source] Expression of FasL in squamous cell carcinomas of the cervix and cervical intraepithelial neoplasia and its role in tumor escape mechanism,CANCER, Issue 5 2006Ramy Ibrahim M.D. Abstract BACKGROUND To date, several mechanisms have been described by which malignant cells escape from the immune system. One of these is through the expression of FasL. The authors hypothesized that the Fas/FasL interaction enables cervical carcinoma cells to induce apoptosis of the cells of the immune system and thereby escape from them. METHODS The authors tested the expression of FASL on the surface of cervical carcinoma tissues. Next, they stained the same cervical tissues with anti-human leukocyte common antigen and TUNEL to identify apoptotic cells. An in vitro functional assay was then done to test if the FASL expressed on the surface of cervical carcinoma cell lines was or was not responsible for inducing apoptosis in T-cells. Finally, they compared the expression of FASL on normal and dysplastic cervical tissues. RESULTS Ninety-four percent of the cervical carcinoma tissues the authors tested expressed FasL and the majority of the apoptotic cells in the specimens were leukocytes with very few tumor cells. In the in vitro functional assay, only the Fasl expressing cell line and not the Fasl negative cell line was able to induce apoptosis of the Fas-expressing Jurkat cells. On examining the normal cervical tissues, the authors found that the expression of Fasl was confined to the basal cell layer with loss of expression observed in the suprabasal layers, which made it an immune privileged site. Conversely, there was persistent expression of FasL in the dysplastic layers in cervical dysplasia and squamous cell carcinoma specimens. CONCLUSIONS The findings of the current study support the authors' hypothesis that persistent expression of FasL plays a role in the ability of cervical carcinoma cells to escape from the immune system. Cancer 2006. Published 2006 by the American Cancer Society. [source] The expression of Bcl-2 family proteins differs between nonsmall cell lung carcinoma subtypesCANCER, Issue 7 2005Helen K. Berrieman Ph.D. Abstract BACKGROUND Proteins of the Bcl-2 family play a key role in the control of apoptosis and carry out both proapoptotic and antiapoptotic functions. However, with the exception of Bcl-2 itself, little is known about the expression of these potentially critical proteins in nonsmall cell lung carcinoma. METHODS Immunohistochemistry was used to study the expression of Bcl-2 and 6 other Bcl-2 family proteins in a pilot series of 41 archival nonsmall cell lung carcinoma specimens (19 adenocarcinomas and 22 squamous cell carcinomas). RESULTS Overexpression of the apoptosis inhibitors Bcl-2 and Bcl-XL was observed in 10 of 41 samples (24%) and in 11 of 41 samples (27%), respectively. Loss of expression of proapoptotic proteins was observed as follows: Bak, 24 of 41 samples (59%); Bad, 21 of 41 samples (51%); Bid, 20 of 41 samples (49%); Bax, 14 of 41 samples (34%); and Bim/Bod, 2 of 41 samples (5%). Statistically significant differences in expression between adenocarcinoma samples and squamous cell carcinoma samples were observed for Bcl-XL (overexpression in 11 of 19 adenocarcinomas [58%] vs. 0 of 22 squamous cell carcinomas [0%]; P < 0.001) and for Bad (loss of expression in 5 of 19 adenocarcinomas [26%] vs. 16 of 22 squamous cell carcinomas [73%]; P = 0.004). CONCLUSIONS Although this was only a pilot study, the results revealed significant differences in the expression of apoptosis-related proteins both between individual samples of nonsmall cell lung carcinoma and between the two main histologic subtypes. Such differences may play a role in the development of lung tumors; and, if it is found that these differences are of clinical importance, then it may be required to regard nonsmall cell lung carcinoma subtypes as separate entities rather than as one disease. Cancer 2005. © 2005 American Cancer Society. [source] Prostate carcinoma tissue proteomics for biomarker discoveryCANCER, Issue 12 2003Yaxin Zheng M.D. Abstract BACKGROUND The advent of the prostate-specific antigen (PSA) test has had a profound impact on the diagnosis and treatment of prostate carcinoma. However, the use of PSA levels alone for screening for prostate carcinoma was compromised by the variations in the amount of PSA produced by the benign prostatic tissue specimens. Proteins were involved in various pathways that determine the behavior of a cell. Therefore, information regarding proteins may reveal drug targets and/or markers for early detection. METHODS The authors used surface-enhanced laser desorption/ionization time-of-flight mass spectrometry to determine the protein profiles from fresh tissues of the prostate. Laser capture microdissection was performed to isolate pure populations of cells. RESULTS The authors identified a protein with an average m/Z of 24,782.56 ± 107.27 that was correlated with the presence of prostate carcinoma. Furthermore, using laser capture microdissection, they demonstrated that the origin of this protein, which the authors designated PCa-24, was derived from the epithelial cells of the prostate. PCa-24 expression was detected in 16 of 17 (94%) prostate carcinoma specimens but not in paired normal cells. In addition, this protein was not expressed in any of the 12 benign prostatic hyperplasia specimens that were assayed. CONCLUSIONS PCa-24 may be useful a marker for prostate carcinoma. Cancer 2003;98:2576,82. © 2003 American Cancer Society. [source] Intra-tumoral interleukin-6 down-regulation system and genetic mutations of tumor suppressor genes in colorectal carcinomaCANCER, Issue 5 2002Chikao Miki M.D. Abstract BACKGROUND The interleukin (IL)-1-IL-6 network, the most potent cascade of pro-inflammatory cytokines, plays an autocrine role in tumor growth. The IL-1-IL-6 network is down-regulated by a phased cytokine inhibitor IL-1 receptor antagonist (ra) and an anti-inflammatory cytokine IL-10. The current study evaluated this down-regulation system in colorectal carcinoma and its relation to the genetic alteration of tumor suppressor genes. METHODS Seventy-four specimens of primary colorectal carcinoma and normal mucosa were collected to measure tissue concentrations of cytokines. Polymerase chain reaction amplification was performed to investigate the loss of heterozygosity of the microsatellite markers on chromosomes 17p and 18q. RESULTS The IL-1ra/IL-6 ratio in the carcinoma specimens was lower than ratios in adenomas and normal mucosae and decreased with disease progression. The IL-1ra/IL-6 ratio in early cancers tended to be lower than that in adenomas and normal mucosae. However, the tissue concentrations of IL-1, and IL-10 were not associated with any clinicopathologic parameters. The tissue IL-1ra concentration correlated with that of IL-6 only in adenomas and early cancers. Immunohistochemically, IL-1ra and IL-6 were localized in the tumor cytoplasm. A reduced tissue IL-1ra/IL-6 ratio in the carcinomas correlated with poor prognosis and was associated with the loss of heterozygosity of the microsatellite markers on chromosomes 18q. CONCLUSIONS There is an IL-6-IL-1ra network system in colorectal tumors, but this system deteriorates with carcinogenesis and tumor growth. The deterioration of this network system was associated with the allelic loss of a portion of chromosome 18q, reflecting the genetic alteration of tumor suppressor genes. Cancer 2002;94:1584,92. © 2002 American Cancer Society. DOI 10.1002/cncr.10324 [source] | ||||||||||||||||||||||