keratan-sulfate and Carcinoma--Squamous-Cell

Resistance to cisplatin is a major obstacle to successful treatment of head and neck squamous cell carcinoma (HNSCC). To investigate the molecular mechanism of this resistance, we compared the gene expression profiles between the cisplatin-sensitive SCC cell lines (Sa-3, H-1 and KB) and the cisplatin-resistant cell lines established from them (Sa-3R, H-1R and KB-R) using Affymetrix U133 Plus 2.0 microarray. We identified 199 genes differentially expressed in each group. To identify important functional networks and ontologies to cisplatin resistance, the 199 genes were analyzed using the Ingenuity Pathway Analysis Tool. Fifty-one of these genes were mapped to genetic networks, and we validated the top-10 upregulated genes by real-time reverse transcriptase-polymerase chain reaction. Five novel genes, LUM, PDE3B, PDGF-C, NRG1 and PKD2, showed excellent concordance with the microarray data. In 48 patients with oral SCC (OSCC), positive immunohistochemical staining for the five genes correlated with chemoresistance to cisplatin-based combination chemotherapy. In addition, the expression of the five genes predicted the patient outcomes with chemotherapy. Furthermore, siRNA-directed suppressed expression of the five genes resulted in enhanced susceptibility to cisplatin-mediated apoptosis. These results suggested that these five novel genes have great potential for predicting the efficacy of cisplatin-based chemotherapy against OSCC. Global gene analysis of cisplatin-resistant cell lines may provide new insights into the mechanisms underlying clinical cisplatin resistance and improve the efficacy of chemotherapy for human HNSCC.

Topics: Aged; Antineoplastic Agents; Blotting, Western; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Survival; Chondroitin Sulfate Proteoglycans; Cisplatin; Cyclic Nucleotide Phosphodiesterases, Type 3; Drug Resistance, Neoplasm; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Genetic Predisposition to Disease; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratan Sulfate; Lumican; Lymphokines; Male; Middle Aged; Neuregulin-1; Platelet-Derived Growth Factor; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; TRPP Cation Channels

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 (SLRP) family and is reported to be overexpressed during the wound healing process of the cornea, and ischemic and reperfused heart. In the carcinomatous tissues, lumican is overexpressed in human breast and pancreatic cancer tissues. In the present study, we aimed to clarify the expression of lumican mRNA and its protein in human cervical cancer cell lines (CaSki, ME-180 and HeLa cells) and their localization in normal and cancerous human cervical tissues. Reverse transcription-polymerase chain reaction and Western blot analysis revealed the expression of lumican mRNA and its protein in CaSki, ME-180 and HeLa cells. No or weak immunoreactivity of the lumican protein was observed in stroma but not in squamous and ductal cells of non-cancerous uterine cervical tissues. In 21 of 28 (75%) cervical cancer cases, the lumican protein was strongly expressed in cancer cells, and accumulated particularly in cancer cells at the periphery of the cancer nests. It was also expressed in the fibroblasts adjacent to the cancer cells. In situ hybridization analysis revealed that lumican mRNA was not expressed in squamous or ductal epithelial cells in non-cancerous tissues, but was expressed in most cancer cells and stromal fibroblasts in uterine cervical cancer tissues. The lumican protein was not localized in normal squamous or ductal cells close to cancer cells, but its mRNA was strongly expressed in the same cells. To our knowledge, this is the first report on lumican synthesized by squamous cell carcinomas. These findings may indicate that the accumulated lumican protein in cancer cells at the periphery of cancer nests may play roles in the growth or invasion of human cervical cancer cells.

Topics: Blotting, Western; Carcinoma, Squamous Cell; Chondroitin Sulfate Proteoglycans; Female; HeLa Cells; Humans; Immunohistochemistry; In Situ Hybridization; Keratan Sulfate; Lumican; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured; Uterine Cervical Neoplasms

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