Human CA9 peptide

Amelotin (AMTN) is an enamel protein that is localized in the basal lamina of ameloblasts in their maturation stage and the internal basal lamina of junctional epithelium (JE) and it is suggested that AMTN could be involved in the dentogingival attachment. To elucidate the transcriptional regulation of human AMTN gene in inflamed gingiva, we have analyzed the effect of tumor necrosis factor-alpha (TNF-alpha) on the expression of AMTN gene in Ca9-22 and Sa3 human gingival epithelial cells. Total RNAs were extracted from Ca9-22 and Sa3 cells after stimulation by TNF-alpha (10 ng/ml). AMTN mRNA and protein levels were measured by real-time PCR and Western blotting. Transient transfection analyses were completed using the various lengths of human AMTN gene promoter constructs with or without TNF-alpha. Gel mobility shift and chromatin immunoprecipitation assays were performed to investigate the transcription factors bindings to the human AMTN gene promoter by TNF-alpha. TNF-alpha (10 ng/ml) increased AMTN mRNA and protein levels after 12 h. TNF-alpha induced luciferase activities of human AMTN gene promoter constructs (- 211AMTN, - 353AMTN, and - 501AMTN). TNF-alpha-induced luciferase activities were partially inhibited in the mutation - 353AMTN constructs that included 3-bp mutations in CCAAT enhancer-binding protein 1 (C/EBP1), C/EBP2 and Ying Yang 1 (YY1) elements. Transcriptional activities induced by TNF-alpha were inhibited by protein kinase A, Src-tyrosine kinase, MEK1/2, p38 kinase, NF-kappa B, and PI3-kinase inhibitors. Gel shift assays showed that TNF-alpha increased nuclear proteins binding to two types of C/EBP elements (C/EBP1 and C/EBP2) and YY1 element. The results of the chromatin immunoprecipitation assays showed that C/EBP beta binding to C/EBP1 and C/EBP2, and YY1 binding to YY1 were increased by TNF-alpha. These findings demonstrated that TNF-alpha stimulates AMTN gene transcription in human gingival epithelial cells via C/EBP1, C/EBP2, and YY1 elements in the human AMTN gene promoter.

This retrospective analysis evaluated the mechanism of bevacizumab plus chemotherapy (BV+CT) for preventing brain metastasis derived from lung cancer. From the total of 159 patients with advanced non-small cell lung cancer (NSCLC), 110 received BV+CT and 49 received CT. After medication, both groups had 15 patients with brain metastases (14 vs 31%, P<0.05). With BV+CT treatment, 40 patients (33.89%) survived, whereas only 11 patients (18.64%) survived with CT treatment. The outcome for the BV+CT group was significantly better than that for the CT group only for vascular endothelial growth factor (VEGF)-positive patients. A post-treatment with BV+CT was significantly reduced than with CT only for patients with high carbonic anhydrase-9 (CA9) expression. This retrospective analysis provides supportive evidence that BVzCT can significantly reduce the incidence of brain metastasis in patients with advanced NSCLC compared with CT alone. Vascular endothelial growth factor - positive patients may benefit more from BV treatment and the outcomes with BV may be related to CA9 expression.