RESUMO
UNLABELLED: Genomic amplification of regional chromosome 8q24 is a common event in human cancers. In hepatocellular carcinoma (HCC), a highly aggressive malignancy that is rapidly fatal, recurrent 8q24 gains can be detected in >50% of cases. In this study, attempts to resolve the 8q24 region by way of array comparative genomic hybridization for affected genes in HCC revealed distinctive gains of block of proliferation 1 (BOP1). Gene expression evaluation in an independent cohort of primary HCC (n = 65) revealed frequent BOP1 up-regulation in tumors compared with adjacent nontumoral liver (84.6%; P < 0.0001). Significant associations could also be drawn between increased expressions of BOP1 and advance HCC staging (P = 0.004), microvascular invasion (P = 0.006), and shorter disease-free survival of patients (P = 0.02). Examination of expression of C-MYC, a well-known oncogene located in proximity to BOP1, in the same series of primary HCC cases did not suggest strong clinicopathologic associations. Functional investigations by small interfering RNA-mediated suppression of BOP1 in HCC cell lines indicated significant inhibition on cell invasion (P < 0.005) and migration (P < 0.05). Overexpression of BOP1 in the immortalized hepatocyte cell line L02 showed increase cellular invasiveness and cell migratory rate (P < 0.0001). In both gene knockdown and ectopic expression assays, BOP1 did not exert an effect on cell viability and proliferation. Evident regression of the epithelial-mesenchymal transition (EMT) phenotype was readily identified in BOP1 knockdown cells, whereas up-regulation of epithelial markers (E-cadherin, cytokeratin 18, and γ-catenin) and down-regulation of mesenchymal markers (fibronectin and vimentin) were seen. A corresponding augmentation of EMT was indicated from the ectopic expression of BOP1 in L02. In addition, BOP1 could stimulate actin stress fiber assembly and RhoA activation. CONCLUSION: Our findings underline an important role for BOP1 in HCC invasiveness and metastasis potentials through inducing EMT and promoting actin cytoskeleton remodeling.
Assuntos
Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/fisiopatologia , Transição Epitelial-Mesenquimal/fisiologia , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/fisiopatologia , Proteínas/fisiologia , Diferenciação Celular/fisiologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células , Estudos de Coortes , Progressão da Doença , Células Epiteliais/patologia , Células Epiteliais/fisiologia , Feminino , Humanos , Fígado/patologia , Fígado/fisiopatologia , Masculino , Mesoderma/patologia , Mesoderma/fisiologia , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-myc/fisiologia , Proteínas de Ligação a RNA , Regulação para Cima/fisiologiaRESUMO
Thymic stromal lymphopoietin (TSLP) is an epithelial cell derived cytokine belonging to the IL-7 family and a key initiator of allergic inflammation. Two main isoforms of TSLP, classified as long- (lfTSLP) and short-form (sfTSLP), have been reported in human, but their expression patterns and role(s) in cancers are not yet clear. mRNA expression was examined by isoform-specific RT-PCR and RNA in situ hybridisation. Epigenetic regulation was investigated by chromatin immunoprecipitation-PCR and bisulfite sequencing. Tumour progression was investigated by gene overexpression, cell viability assay, cancer organoid culture and transwell invasion. Signals were investigated by proteome profiler protein array and RNA-sequencing. With the use of isoform-specific primers and probes, we uncovered that only sfTSLP was expressed in the cell lines and tumour tissues of human ovarian and endometrial cancers. We also showed the epigenetic regulation of sfTSLP: sfTSLP transcription was regulated by histone acetylation at promoters in ovarian cancer cells, whereas silencing of the sfTSLP transcripts was regulated by promoter DNA methylation in endometrial cancer cells. In vitro study showed that ectopically overexpressing sfTSLP promoted tumour growth but not invasion. Human phosphokinase array application demonstrated that the sfTSLP overexpression activated phosphorylation of multiple intracellular kinases (including GSK3α/ß, AMPKα1, p53, AKT1/2, ERK1/2 and Src) in ovarian cancer cells in a context-dependent manner. We further investigated the impact of sfTSLP overexpression on transcriptome by RNA-sequencing and found that EFNB2 and PBX1 were downregulated in ovarian and endometrial cancer cells, suggesting their role in sfTSLP-mediated tumour growth. In conclusion, sfTSLP is predominantly expressed in ovarian and endometrial cancers and promotes tumour growth.
RESUMO
We have examined expression of L1 and the polysialic acid-associated form of the neural cell adhesion molecule (PSA-NCAM) in mouse embryos during the major period of axon growth in the retinofugal pathway to determine whether they are expressed in patterns that relate to the changes in axon organization in the pathway. Immunostaining for L1 and PSA-NCAM was found on all axons in the retina and the optic stalk. In the chiasm, while L1 immunoreactivity remained high on the axons, PSA-NCAM staining was obviously reduced. At the threshold of the optic tract, L1 immunoreactivity was maintained only in a subpopulation of axons, whereas PSA-NCAM staining was dramatically elevated in axons at the caudal part of the tract. Further investigations of the tract showed that both L1 and PSA-NCAM were preferentially expressed on the dorsal but not ventral optic axons, indicating a regionally specific change of both adhesion molecules on the axons at the chiasm-tract junction. Moreover, intense PSA-NCAM expression was also observed in the tract of postoptic commissure (TPOC), which lies immediately caudal to the optic tract. Immunohistochemical and retrograde tracing studies showed that these PSA-NCAM-positive axons arose from a population of cells rostral to the CD44-positive chiasmatic neurons. These findings indicate that, in addition to the chiasmatic neurons, these PSA-NCAM-positive diencephalic cells also contribute axons to the TPOC. These early generated commissural axons together with the regionally specific pattern of cell adhesion molecule expression on the optic axons may control formation of the partial retinotopic axon order in the optic tract through homophilic or heterophilic interactions that involve PSA-NCAM.