Bioinformatics screening of gene expression profile and diagnostic application of meningeal carcinoma.

OBJECTIVE: The aim of this study was to research gene expression profiles and diagnostic applications of meningeal carcinoma based on bioinformatics. MATERIALS AND METHODS: We used the Gene Expression Omnibus (GEO) database to obtain the GSE43290 dataset based on the expression data of normal meninges and meningiomas consisting of 51 samples divided into two groups (47 samples of meningioma tumors and four samples of normal meninges). We used the GEO2R tool to identify differentially expressed genes (DEGs) by setting the log2 fold change as greater than two and an adjusted p-value lower than 0.05. We used the database for annotation, visualization and integrated discovery (DAVID) to perform gene ontology, biological pathways and functional annotation of the DEGs. A search Tool for the Retrieval of Interacting Gene database (STRING) was used to obtain Protein-Protein Interaction (PPI) and modular networks based on the Markov clustering algorithm. RESULTS: Our study identified 358 significant DEGs, of which 343 were downregulated genes while 15 were upregulated. Five significant hub genes (CXCL8, AGT, CXCR4, CXCL12 and CXCL2) were associated with various biological pathways, molecular functions and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The DEGs were enriched in biological pathways of chemokine-mediated signaling, positive regulation of leukocyte chemotaxis, second messenger-mediated signaling, induction of positive chemotaxis, CXCR chemokine receptor binding and activities of cytokines. CONCLUSIONS: These hub genes and pathways could be targeted in clinical research to discover new treatments for meningeal carcinoma.

Effect of lncRNA LET on proliferation and invasion of osteosarcoma cells.

OBJECTIVE: To investigate the expression of long non-coding RNA (LncRNA) LET in osteosarcoma and its effect on the proliferation, apoptosis, migration and invasion of osteosarcoma cells. PATIENTS AND METHODS: The expression of lncRNA LET was detected in osteosarcoma tissues and cell lines (MG63 and hFOB1.19). MG63 cells stably overexpressing lncRNA LET were constructed by lentiviral. The effects of lncRNA LET overexpression on the proliferation, apoptosis, migration and invasion of osteosarcoma cells were detected by cell counting kit-8 (CCK-8), flow cytometry and transwell chamber assay. RESULTS: The expression of lncRNA LET in osteosarcoma tissues and MG63 cells was significantly down-regulated. Overexpression of lncRNA LET significantly inhibited the proliferation, migration, invasion, and induced apoptosis of MG63 cells. CONCLUSIONS: LncRNA LET was participated in the development of osteosarcoma, and may be used as a potential molecular target for the treatment of osteosarcoma.

MiR-361 inhibits osteosarcoma cell lines invasion and proliferation by targeting FKBP14.

OBJECTIVE: MicroRNAs have been reported to contribute to the development of osteosarcoma via negatively regulating the target genes. Nevertheless, the undiscovered function of miR-361 on osteosarcoma development remains uncertain. PATIENTS AND METHODS: MiR-361 and FKBP14 (FK506-binding protein 14) expression in osteosarcoma samples were detected by Real-time polymerase chain reaction (PCR). Cells invasive ability was examined via the transwell invasion assay. The luciferase reporter assay was used to examine the regulation mechanism. The protein level of FKBP14 was detected by Western blot. Cell Counting Kit-8 (CCK-8) was used to detect cell lines proliferation. RESULTS: MiR-361 was reduced both in osteosarcoma samples and cell lines. Up-regulation of miR-361 significantly inhibited cells invasive and proliferative abilities, while down-regulation of miR-361 promoted cell lines invasion and proliferation. miR-361 could negatively regulate FKBP14 in osteosarcoma. Suppression of FKBP14 could reverse the function of miR-361 inhibitor. CONCLUSIONS: MiR-361 inhibits osteosarcoma cell lines invasion and proliferation by targeting FKBP14.

Pivotal role of microRNA-9 in osteosarcoma tumorigenesis and tumor progression.

MicroRNA-9 (miR-9) has a well-established role in various tumors; the clinical significance and potential mechanism of miR-9 in human osteosarcoma (OS) has not been elucidated. The aim of this study was to investigate the mechanism and role of miR-9 expression in osteosarcoma cells. miR-9 expression in the OS cell line MG-63 and OS tissues was compared to that in a human osteoblastic cell line (hFOB 1.19) and adjacent normal tissues, respectively, by reverse transcriptase-polymerase chain reaction. miR-9 expression was downregulated by introducing small interfering RNA against miR-9 (si-miR-9) into the cells, and the proliferative, migratory, and invasive capacities of si-miR-9-transfected MG-63 cells were compared to those of control MG-63 cells. miR-9 was significantly upregulated in OS tissues and cell lines compared to the corresponding non-cancerous bone tissues (P < 0.05) and human osteoblastic cell line (P < 0.05), respectively. Upregulated miR-9 expression was also associated with increased cell proliferation (P < 0.05), migration (P < 0.05), and invasion (P < 0.05), and decreased apoptotic ability (P < 0.05). These results suggest that miR-9 may play a pivotal role in tumorigenesis and tumor progression in osteosarcoma.

Parallel proteomic analysis in muscle-invasive bladder transitional cell carcinoma and cancer-related stroma.

To bring about improvements in cancer biology research and elucidate mechanism-based therapeutic targets, we studied the proteome expression profile of purified normal urothelial cells (cancer cells) and normal stromal cells (cancerous stromal cells). Based on the expression profile, biomarker discovery and the mechanisms of multi-step carcinogenesis were explored. We found that 1412/1403 unique proteins commonly appeared in 4 sets of paired cancer/normal tissue, and 1753 proteins were differentially expressed. Three hundred and forty-one proteins were repeatedly expressed in both cancer and cancer stromal cells; 358 proteins were repeatedly expressed in both normal urothelial and normal stromal cells. Among them, 186/203 proteins were specific repeat expressions in cancer/normal tissue and thought to play an important role in cancer-stroma interactions. Differential proteins were further analyzed using bioinformatic tools and compared with the published literature. GO enrichment/depletion analysis indicated that carcinogenesis involved all the biological processes and all the cellular components. Five hundred and sixty-eight differential proteins were located in the well-known biological Kyoto Encyclopedia of Genes and Genomes pathways, including metabolic pathways, ribosome spliceosome, and endocytosis. One hundred and thirty-nine of the 186 proteins that displayed specific repeat expressions in cancer tissue were located in the biological Kyoto Encyclopedia of Genes and Genomes pathways and are thought to be candidate biomarkers for targeted therapy.

Human catalytic antibodies with glutathione peroxidase activity.

In order to generate catalytic antibodies with glutathione peroxidase (GPx) activity, we prepared GSH-S-DNP butyl ester and GSH-S-DNP benzyl ester as the haptens. Two ScFvs that bound specifically to the haptens were selected from the human phage-displayed antibody library. The two ScFv genes were highly homologous, consisting of 786 bps and belonging to the same VH family-DP25. In the premise of maintaining the amino acid sequence, mutated plasmids were constructed by use of the mutated primers in PCR, and they were over-expressed in E. coli. After the active site serine was converted into selenocysteine with the chemical modifying method, we obtained two human catalytic antibodies with GPx activity of 72.2U/micromol and 28.8U/micromol, respectively. With the aid of computer mimicking, it can be assumed that the antibodies can form dimers and the mutated selenocysteine residue is located in the binding site. Furthermore, the same Ping-Pong mechanism as the natural GPx was observed when the kinetic behavior of the antibody with the higher activity was studied.