keratan-sulfate and Adenocarcinoma

Keratan sulfate (KS) is a sulfated linear polymer of N-acetyllactosamine. Proteoglycans carrying keratan sulfate epitopes were majorly observed in cornea, cartilage and brain; and mainly involved in embryonic development, cornea transparency, and wound healing process. Recently, expression of KS in cancer has been shown to be highly associated with advanced tumor grade and poor prognosis. Therefore, we aimed to identify the expression of KS epitope in human pancreatic cancer primary and metastatic tumor lesions. Immunohistochemical analysis of KS expression was performed on primary pancreatic tumors and metastatic tissues. We observed an increased expression of KS epitope on primary tumor tissues compared to uninvolved normal and tumor stroma; and is associated with worse overall survival. Moreover, lung metastatic tumors show a higher-level expression of KS compared to primary tumors. Interestingly, KS biosynthesis specific glycosyltransferases expression was differentially regulated in metastatic pancreatic tumors. Taken together, these results indicate that aberrant expression of KS is predictive of pancreatic cancer progression and metastasis and may serve as a novel prognostic biomarker for pancreatic cancer.

Topics: Adenocarcinoma; Biomarkers, Tumor; Female; Follow-Up Studies; Gene Expression Profiling; Humans; Keratan Sulfate; Male; Middle Aged; Pancreatic Neoplasms; Prognosis; Survival Rate; Tissue Array Analysis; Tumor Cells, Cultured

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

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

Chronic pancreatitis is a chronic inflammatory disorder of the pancreas. The etiology is multi-fold, but all lead to progressive scarring and loss of pancreatic function. Early diagnosis is difficult; and the understanding of the molecular events that underlie this progressive disease is limited. In this study, we investigated differential proteins associated with mild and severe chronic pancreatitis in comparison with normal pancreas and pancreatic cancer. Paraffin-embedded formalin-fixed tissues from five well-characterized specimens each of normal pancreas (NL), mild chronic pancreatitis (MCP), severe chronic pancreatitis (SCP) and pancreatic ductal adenocarcinoma (PDAC) were subjected to proteomic analysis using a "label-free" comparative approach. Our results show that the numbers of differential proteins increase substantially with the disease severity, from mild to severe chronic pancreatitis, while the number of dysregulated proteins is highest in pancreatic adenocarcinoma. Important functional groups and biological processes associated with chronic pancreatitis and cancer include acinar cell secretory proteins, pancreatic fibrosis/stellate cell activation, glycoproteins, and inflammatory proteins. Three differential proteins were selected for verification by immunohistochemistry, including collagen 14A1, lumican and versican. Further canonical pathway analysis revealed that acute phase response signal, prothrombin activation pathway, and pancreatic fibrosis/pancreatic stellate cell activation pathway were the most significant pathways involved in chronic pancreatitis, while pathways relating to metabolism were the most significant pathways in pancreatic adenocarcinoma. Our study reveals a group of differentially expressed proteins and the related pathways that may shed light on the pathogenesis of chronic pancreatitis and the common molecular events associated with chronic pancreatitis and pancreatic adenocarcinoma.

Topics: Acinar Cells; Adenocarcinoma; Biological Specimen Banks; Chondroitin Sulfate Proteoglycans; Cluster Analysis; Collagen; Extracellular Matrix Proteins; Glycoproteins; Humans; Immunohistochemistry; Keratan Sulfate; Lumican; Pancreas; Pancreatic Neoplasms; Pancreatic Stellate Cells; Pancreatitis, Chronic; Proteins; Proteomics; Reproducibility of Results; Signal Transduction; Staining and Labeling; Versicans

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

Lumican is a member of a small leucine-rich proteoglycan family and its overexpression has been reported in carcinoid tumor, breast, colorectal, neuroendocrine cell, uterine cervical and pancreatic cancers. The expression of lumican in stromal tissues in breast cancer is associated with a high tumor grade, a low estrogen receptor expression level and young age. Lumican expression in the cytoplasm in advanced colorectal cancer is correlated with a poor prognosis. Lumican expression was previously reported in pancreatic cancer, but the role of lumican in pancreatic cancer is still not well understood. In this study, we aimed to clarify the role of lumican in pancreatic cancer. Reverse-transcription polymerase chain reaction and Western blot analyses revealed lumican mRNA and protein expression in six pancreatic ductal adenocarcinoma cell lines (i.e. PANC-1, MIA PaCa-2, KLM-1, Capan-1, PK-1 and PK-8). On the basis of its immunoreactivity, lumican was found to be localized in islet cells of normal pancreatic tissues, but not in exocrine cells. In pancreatic cancer tissues, lumican was predominantly localized in the cytoplasm of cancer cells in 30 out of 53 (56.6%) cancer patients, whereas lumican was detected in stromal tissues in 36 out of 53 (67.9%) cancer patients. Lumican expression in pancreatic cancer cells did not correlate with clinicopathological factors, whereas lumican expression in stromal tissues correlated with the female gender, advanced stage, retroperitoneal and duodenal invasion and residual tumor (p=0.030, 0.038, 0.049, 0.049 and 0.048, respectively). Patients with lumican-positive cancer cells tended to survive longer than those with lumican-negative cancer cells (p=0.286), but patients with lumican-positive stromal tissues had shorter survival than those with lumican-negative stromal tissues (p=0.062). These results suggest that lumican in stromal tissues plays an important role in the growth and invasion of pancreatic cancer.

Topics: Adenocarcinoma; Carcinoma, Pancreatic Ductal; Cell Line, Tumor; Chondroitin Sulfate Proteoglycans; Gene Expression Regulation, Neoplastic; Humans; Islets of Langerhans; Keratan Sulfate; Lumican; Lymphatic Metastasis; Neoplasm Invasiveness; Pancreas; Pancreatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stromal Cells

Lumican is a member of a small leucine-rich proteoglycan family, and it is reportedly overexpressed in human breast cancer. The expression of lumican in the extracellular matrix in breast cancer is associated with a high tumor grade, low estrogen receptor levels and young age. Lumican expression has been previously reported in colorectal cancer, but the role of lumican in the tumor is not well understood. In this study, we examined the expression and role of lumican in advanced colorectal cancer. Immunohistochemical staining was performed on 158 patients who underwent curative surgery for advanced colorectal cancer with lymph node metastasis. In the normal colorectal tissues, lumican immunoreactivity was observed in the fibroblasts and neural cells, but not in the colorectal epithelial cells. Lumican was localized in the cytoplasm of the cancer cells and its overexpression was detected in 99 of the 158 (62.7%) colorectal cancer patients. Clinicopathologically, there was no association of lumican expression with age, sex, histological typing, or venous and lymphatic invasion. However, lumican expression tended to correlate with the spread of lymph node metastasis and the depth of tumor invasion (p=0.136 and 0.135, respectively). Furthermore, the survival rate was significantly lower in patients with a high lumican expression level than in those with a low lumican expression level (p=0.048). These results indicate that lumican expression is a potential prognostic factor in patients with advanced colorectal cancer with nodal metastasis.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Chondroitin Sulfate Proteoglycans; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Keratan Sulfate; Lumican; Lymphatic Metastasis; Male; Middle Aged; Prognosis; Survival Analysis; Survival Rate

The malignant behavior of cancers depends on the microenvironmental context. We investigated compositional alterations of the extracellular matrix (ECM) in pancreatic cancer, with special emphasis on the proteoglycans decorin, lumican, and versican. Compared with normal controls (n=18), marked overexpression of these proteoglycans was observed in pancreatic cancer tissues (n=30) by quantitative RT-PCR (p<0.0001). Immunohistochemistry revealed abundance of proteoglycans in the ECM of pancreatic cancer specimens, whereas tumor cells themselves were devoid of either decorin, lumican or versican. RT-PCR confirmed pancreatic stellate cells (PSCs) as the major source of these proteins. Interestingly, TGFbeta1 and conditioned medium derived from pancreatic cancer cell lines synergistically suppressed the expression of known anti-tumor factors decorin and lumican, but stimulated the expression of pro-metastatic factor versican in cultured PSCs. These findings indicate that malignant cells can actively influence the composition of the ECM through TGFbeta1 and other soluble factors, altering their microenvironment in a tumor-favorable way.

Topics: Adenocarcinoma; Adult; Aged; Chondroitin Sulfate Proteoglycans; Decorin; Extracellular Matrix Proteins; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratan Sulfate; Lectins, C-Type; Lumican; Male; Middle Aged; Pancreatic Neoplasms; Proteoglycans; Reverse Transcriptase Polymerase Chain Reaction; Tumor Cells, Cultured; Versicans

Immunohistochemical staining was performed on normal, endometriotic, neoplastic endometrial tissues and various epithelial tissues of other organs using a monoclonal antibody, 5D4, which was raised originally against skeletal and corneal keratan sulfate. The results showed that this antibody exhibited strong and consistent reactivity to glandular epithelia found in normal and endometriotic tissues. In endometrial carcinomas, moderate reactivity was noted in well- to moderately differentiated adenocarcinoma. Other epithelial tissues examined showed almost no or faint reactivities. These results suggested that 5D4 would be a useful tool for identifying the endometrial glandular epithelia in histochemical studies.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Endometrial Neoplasms; Endometriosis; Endometrium; Female; Humans; Immunoenzyme Techniques; Keratan Sulfate; Uterine Neoplasms