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Purpose To explore the association of NET-1 expression with the angiogenesis of cervical squamous cell carcinoma and the related molecular mechanism.Methods Immunohistochemistry MaxVision was used to detect the expression of NET-1,NF-κB p65 and VEGF in 80 cases of cervical squamous cancer tissue and 10 cases of chronic cervicitis tissue.The microvascular density (MVD) was assessed by labeling endothelia with CD34-antibody and counted according to Weinner's standard.The eukaryotic expression plasmid pCMV6-NET-1 was transfected into cervical squamous cancer cells by cationic liposome in order to obtain over expression of NET-1 gene.In order to inhibit the activation of NF-kappa B,cervical squamous carcinoma cells were treated with NF-kappa B specific inhibitor PDTC.The protein expression in SiHa cells was assessed by Western blotting.Results The expression level of NET-1 in the cervical squamous carcinoma tissue was significantly higher than that in the tissue of control group.NET-1 expression was positively correlated to the MVD,NF-κB p65 and VEGF expression,respectively.Transient transfection of NET-1 gene significantly increased the nuclear expression of NF-κB p65 and cytoplasm expression of VEGF in the SiHa cells.However,pretreatment of SiHa cells with NF-inhibitor PDTC significantly inhibited NET-1 induced upregulation of VEGF expression.Conclusion NET-1 promotes the angiogenesis of cervical squamous carcinoma through the activation of the NF-κB signaling which upregulate the expression of VEGF in the cancer cells.
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Objective To study the effect of neuroepithelial cell transforming gene 1 (Net1) on the cellular radiosensitivity and underlying mechanism.Methods Real-time quantitative PCR was used to measure the variations in Net1 expression level upon irradiation.Radiosensitivity was analyzed by colonyforming assay after Net1-siRNAs.Net1-associated proteins were identified by co-immunoprecipitation.Results The Net1 mRNA level in the cells was increased significantly (t =-10.52,P ＜ 0.05) after irradiation.Compared to the control group,siRNA-mediated silencing of Net1 enhanced cell radiosensitivity (t =15.31,11.65,P ＜0.05).Net1 was found to interact with Ku70,Ku80 and DNA-PKcs under either normal conditions or after irradiation.Conclusions Net1 could protect cells from irradiation by interaction with DNA repair proteins in non-homologous end joining pathway.
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Objective To investigate the correlation between microvessel density (MVD) and expression of NET-1 protein in bladder carcinoma and its prognostic significance. Methods The expression NET-1 protein and MVD marked with CD_ 105 in 43 cases of bladder carcinoma and 10 normal bladder tissues were measured by means of immunohistochemistry SP.The association between them and the relation ship with WHO pathology classification,TNM stage and prognosis were studied. Results MVD were correlated with WHO pathology classification,TNM stage and prognosis (F_1=17.01,F_2=12.06,P
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Objective:To investigate the expression of latent membrane protein-1(LMP-1),NET-1,and vascular endothelial growth factor(VEGF)in non-keratin nasopharyngeal carcinoma(NK-NPC)and its clinical significance.Methods :Sixty biopsy specimens from pathologically-confirmed NK-NPC patients,who were treated in the Affilitated Hospital of Nantong University from May 1999 to May 2003,were included in the present study.Using immunohistochemical techniques (Envison two-step),we examined the expression of LMP-1,NET-1 and VEGF protein in the specimens.The relationship between their expression and elnicopathological parameters and the prognosis was anayzed.Ten specimens of chronic nasopharyngitis served as control.Results:(1)The positive rates of LMP-1,NET-1 and VEGF in NK-NPC were significantly higher than those in the chronic nasopharyngitis(P0.05).(3)NET-1 and VEGF expression in NK-NPC specimens were positively correlated with lymph node metastasis and distant metastasis(P
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To identify new bio-markers as well as potential targets for new drugs for epithelial ovarian cancer (EOC), we compared the gene expression profiles of cancer tissues from 25 EOCs with human ovarian surface epithelium (HOSE) using in-house cDNA microarray specified to EOC. Based on a comprehensive method and information from the National Cancer Institute (NCI) Cancer Genome Anatomy Project (CGAP), the cDNA library was constructed. After excluding the overlapping clones, 768 spots were included in the array. We identified the genes and expressed sequence tags (ESTs) (30 up-regulated and 34 down-regulated) that are differentially expressed in EOC tissues. To confirm the expression data, we performed real time RT-PCR experiments. Using microdissected EOC tissues and cell lines, we investigated the expression status of the NET-1 gene, clusterin gene, and actin-binding LIM protein 1. The information provided here will be useful for identifying genes whose products might serve as molecular signatures for the biomakers of EOCs.