keratan-sulfate and Lung-Neoplasms

Adenocarcinoma (AdC) is the most common lung cancer subtype and is often associated with pleural effusion (PE). Its poor prognosis is attributable to diagnostic delay and lack of effective treatments and there is a pressing need in discovering new biomarkers for early diagnosis or targeted therapies. To date, little is known about lung AdC proteome. We investigated protein expression of lung AdC in PE using the isobaric Tags for Relative and Absolute Quantification (iTRAQ) approach to identify possible novel diagnostic/prognostic biomarkers. This provided the identification of 109 of lung AdC-related proteins. We further analyzed lumican, one of the overexpressed proteins, in 88 resected lung AdCs and in 23 malignant PE cell-blocks (13 lung AdCs and 10 non-lung cancers) using immunohistochemistry. In AdC surgical samples, lumican expression was low in cancer cells, whereas it was strong and diffuse in the stroma surrounding the tumor. However, lumican expression was not associated with tumor grade, stage, and vascular/pleural invasion. None of the lung cancer cell-blocks showed lumican immunoreaction, whereas those of all the other tumors were strongly positive. Finally, immunoblotting analysis showed lumican expression in both cell lysate and conditioned medium of a fibroblast culture but not in those of A549 lung cancer cell line. PE is a valid source of information for proteomic analysis without many of the restrictions of plasma. The high lumican levels characterizing AdC PEs are probably due to its release by the fibroblasts surrounding the tumor. Despite the role of lumican in lung AdC is still elusive, it could be of diagnostic value.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Cell Line, Tumor; Chondroitin Sulfate Proteoglycans; Female; Humans; Immunohistochemistry; Keratan Sulfate; Lumican; Lung Neoplasms; Male; Neoplasm Grading; Neoplasm Staging; Pleural Effusion, Malignant; Proteome; Proteomics

Giant cell tumor of bone (GCTB) displays worrisome clinical features such as local recurrence and occasionally metastatic disease which are unpredictable by morphology. Additional routinely usable biomarkers do not exist. Gene expression profiles of six clinically defined groups of GCTB and one group of aneurysmal bone cyst (ABC) were determined by microarray (n = 33). The most promising differentially expressed genes were validated by Q-PCR as potential biomarkers in a larger patient group (n = 41). Corresponding protein expression was confirmed by immunohistochemistry. Unsupervised hierarchical clustering reveals a metastatic GCTB cluster, a heterogeneous, non-metastatic GCTB cluster, and a primary ABC cluster. Balanced score testing indicates that lumican (LUM) and decorin (DCN) are the most promising biomarkers as they have lower level of expression in the metastatic group. Expression of dermatopontin (DPT) was significantly lower in recurrent tumors. Validation of the results was performed by paired and unpaired t test in primary GCTB and corresponding metastases, which proved that the differential expression of LUM and DCN is tumor specific rather than location specific. Our findings show that several genes related to extracellular matrix integrity (LUM, DCN, and DPT) are differentially expressed and may serve as biomarkers for metastatic and recurrent GCTB.

Topics: Adolescent; Adult; Biomarkers, Tumor; Bone Neoplasms; Child; Chondroitin Sulfate Proteoglycans; Cluster Analysis; Decorin; Down-Regulation; Extracellular Matrix; Extracellular Matrix Proteins; Female; Gene Expression Profiling; Giant Cell Tumor of Bone; Humans; Immunohistochemistry; Keratan Sulfate; Lumican; Lung Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Oligonucleotide Array Sequence Analysis; Real-Time Polymerase Chain Reaction; Transcriptome; Young Adult

To investigate the effect of the lumican gene on the proliferation of lung adenocarcinoma cell A549 and the possible mechanism.. The lung adenocarcinoma cell line A549 was infected with the recombinant lentivirus carrying the human lumican gene and a stable cell line was obtained via puromycin selection. The expression of lumican mRNA and protein was confirmed by fluorescence quantitative PCR(FQ-PCR) and Western blotting. We examined the cell growth curve, doubling time, proliferation index using MTT assay and flow cytometry and observed the status of the tumor in the experimental animals. Meanwhile, Western blotting was used to detect the expressions of RhoC and p-Akt proteins.. The A549 cell line with the lumican gene over-expression was successfully built. Compared with the control group and the empty vector group, the cells in the experimental group grew faster (P<0.05), the doubling time was shortened (P<0.05), the proliferation index went up (P<0.05), subcutaneous tumors in volume and mass increased and cells of the mitotic figures became more (P<0.05), and the expressions of RhoC and p-Akt proteins were raised (P<0.05). However, there was no obvious difference between the control group and the empty vecor group (P>0.05).. Lumican gene can promote the proliferation of lung adenocarcinoma cell A549, and its mechanism may be related to increased RhoC and p-Akt protein expressions.

Topics: Adenocarcinoma; Animals; Cell Line, Tumor; Cell Proliferation; Chondroitin Sulfate Proteoglycans; Gene Expression; Genetic Vectors; Humans; Keratan Sulfate; Lentivirus; Lumican; Lung Neoplasms; Mice; Proto-Oncogene Proteins c-akt; rho GTP-Binding Proteins; Transduction, Genetic; Transplantation, Heterologous; Tumor Burden

The authors previously reported the expression of keratan sulfate (KS), a glycosaminoglycan, in the epithelium of normal and neoplastic endometria. The aim of this study was to evaluate its potential use as a diagnostic marker, and the expression of KS was investigated in other human epithelial tissues.. Expression was examined immunohistochemically using 102 samples of normal epithelia and 110 samples of carcinomas from the female genital tract (FGT; cervix, endometrium, ovary, fallopian tube), digestive organs (gastrointestinal tract, pancreas, liver), urinary tract, lung, mammary gland, thyroid and mesothelium.. In normal tissues, KS was consistently detected in the FGT and ectopic endometrium (25/26), but was not found in the digestive organs (1/42) and urinary tract (0/6), and was only partly detected in the lung (7/10), mammary gland (3/9) and thyroid (4/4). In malignant tissues, KS was consistently observed in carcinomas of the endometrium, ovary and fallopian tube (29/32), and was partly detected in carcinomas of the lung, mammary gland, thyroid, pancreas and mesothelium, but was absent in carcinomas of the gastrointestinal tract (0/17), liver (0/5) and urinary tract (0/11). Among carcinomas of the FGT, digestive organs and urinary tract, KS positivity suggested the possibility of FGT carcinomas, with 79.5% (31/39) sensitivity and 92.9% (39/42) specificity.. KS is a potentially useful marker for the supportive diagnosis of the primary site of metastatic carcinomas or unknown primary carcinomas, especially in the abdominal cavity.

Topics: Biomarkers, Tumor; Digestive System Neoplasms; Female; Genital Neoplasms, Female; Humans; Immunohistochemistry; Keratan Sulfate; Lung Neoplasms; Urologic Neoplasms

Lumican is a small leucine-rich proteoglycan (SLRP) of the extracellular matrix (ECM) involved in the control of melanoma growth and invasion. The aim of the present study was to analyse the role of lumican in the regulation of the development of lung metastasis.. B16F1 melanoma cells stably transfected with lumican expressing plasmid (Lum-B16F1) were injected to syngenic mice. The lung metastasis was compared to mice injected with mock-transfected B16F1 cells (Mock-B16F1). The expression of lumican, cyclin D1, apoptotic markers, vascular endothelium growth factor (VEGF) and Von Willebrand Factor (vWF) within lung metastasis nodules was investigated by immunohistochemistry. In parallel, cells cultured in presence of lumican were assayed for apoptosis and motility.. We observed that the number and the size of lung metastasis nodules were significantly decreased in mice injected with Lum-B16F1 cells in comparison to Mock-B16F1 cells. This was associated with an increase of tumour cell apoptosis within metastasis nodules but the cell proliferation rate remained constant in the two mice groups. In contrast, the VEGF immunostaining and the number of blood vessels within the lung metastasis nodules were decreased in the lumican-expressing tumours. In vitro, a significant decrease of apoptotic markers in wild type B16F1 cells incubated with increasing amounts of lumican core protein was observed. In addition, pseudotubes formation on Matrigel(R) and the migratory capacity of endothelial cells was inhibited by lumican. Altogether, our results indicate that lumican decreases lung metastasis development not only by inducing tumour cell apoptosis but also by inhibiting angiogenesis.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Apoptosis; Blotting, Western; Cell Movement; Cells, Cultured; Chondroitin Sulfate Proteoglycans; Endothelial Cells; Female; Humans; Image Processing, Computer-Assisted; Keratan Sulfate; Lumican; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Recombinant Proteins; Vascular Endothelial Growth Factor A

Lumican is a member of a small leucine-rich proteoglycan family and is highly expressed in several types of cancer cells and/or stromal tissue. Lumican expression in the cytoplasm in advanced colorectal cancer correlates with poor patient prognosis. The expression of lumican in stromal tissues is associated with a high tumor grade, a low estrogen receptor expression level, and young age in breast cancer and is associated with tumor invasion and advanced stage in pancreatic cancer. In this study, we examined the expression and role of lumican in lung cancer including adenocarcinoma (ADC) and squamous cell carcinoma (SqCC). Immunohistochemically, lumican was weakly expressed in vascular smooth muscle cells, perivascular and peribronchial connective tissues and bronchial epithelium of normal lung tissues. In lung cancer tissues, lumican was localized in the cytoplasm of cancer cells and/or stromal tissues adjacent to cancer cells. In ADC, the expression level of lumican in cancer cells correlated with pleural invasion and larger tumor size, but that of lumican in stromal tissues did not correlate with clinicopathological factors. In SqCC, the expression level of lumican in cancer cells correlated with formation of a keratinized pattern, and stromal lumican expression correlated with vascular invasion. In SqCC and ADC, the expression level of lumican in cancer cells did not correlate with patient prognosis. In lung cancer cell lines, lumican mRNA and protein were expressed in LC-1/Sq and EBC-1 cells established from SqCC, and A549, RERF-LC-KJ and PC-3 cells from ADC. The molecular weight of lumican extracted from the cytoplasm of lung cancer cells differed from that in the culture medium owing to glycosylation of the protein. These findings suggest that the expression pattern and the glycosylated type of lumican in cells and stromal tissues correlate with the aggressiveness of lung SqCC and ADC.

Topics: Adenocarcinoma; Aged; Blotting, Western; Carcinoma, Squamous Cell; Cell Line, Tumor; Chondroitin Sulfate Proteoglycans; Cytoplasm; Female; Gene Expression Regulation, Neoplastic; Glycosylation; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Keratan Sulfate; Lumican; Lung Neoplasms; Male; Middle Aged; Molecular Weight; Neoplasm Invasiveness; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stromal Cells; Time Factors; Treatment Outcome

Topics: Carcinoma, Squamous Cell; Dermatan Sulfate; Diabetes Mellitus, Type 1; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Keratan Sulfate; Lung Neoplasms; Male; Middle Aged; Protein Binding

We have developed a new ELISA to quantify hyaluronate. This ELISA takes advantage of an anti-keratan sulfate antibody to differentiate between the coated aggregating rat chondrosarcoma proteoglycan which captures the hyaluronate and the keratan sulfate-bearing aggregating proteoglycan added subsequently. Absorbance in this assay was shown to be linear to the logarithmic concentration of hyaluronate in the range of 15 to 1000 ng/ml. Pre-treatment of hyaluronate with papain or protease did not interfere with its quantification; in contrast, pre-treatment with 0.1N NaOH significantly interferes with the subsequent measurement of the hyaluronate molecules. The size of the hyaluronate molecule was found to be an important factor in quantification: only large size hyaluronate molecules could be quantified accurately. The ELISA was used to show that human lung carcinomas contain 2 to 500 times as much hyaluronate as normal lung tissue from the same patient.

Topics: Antibodies, Monoclonal; Carcinoma; Chondroitin Sulfate Proteoglycans; Chromatography, Gel; Enzyme-Linked Immunosorbent Assay; Humans; Hyaluronic Acid; Hydrogen-Ion Concentration; Keratan Sulfate; Lumican; Lung; Lung Neoplasms; Molecular Weight