ABSTRACT
BACKGROUND: Angiogenesis is a critical biological process essential for solid cancer growth and metastasis. It has been shown that microRNAs (miRNAs) play a vital role in a variety of biological processes in cancers. However, whether miR-130b is involved in prostate cancer angiogenesis remains ill-defined. METHODS: We performed the miRNA microarray to analyze miRNA expression in human prostate cancer specimens. In vitro gain-of-function assays and loss-of-function assays were conducted to explore the potential functions of miR-130b in human prostate cancer cells. Correlation analysis and dual-luciferase reporter assay were performed to validate whether tumor necrosis factor-α (TNF-α) was a direct target of miR-130b. The Matrigel plug and tumor vascular imaging assays were performed to confirm the anti-angiogenic activity of miR-130b in nude mice. RESULTS: We found that miR-130b was one of the miRNAs being most significantly downregulated. Subsequently, we found that miR-130b expression was markedly downregulated in human prostate cancer cell lines. Down-regulation of miR-130b in prostate cancer cells significantly promoted the proliferation, invasion and tubule formation of human umbilical vein endothelial cells (HUVECs), while ectopic expression of miR-130b blocked prostate cancer angiogenesis in vitro and in vivo. Mechanistic analyses indicated that tumor necrosis factor-α (TNF-α) was regulated by miR-130b directly. MiR-130b attenuated nuclear factor-κB (NF-κB) signaling and its downstream gene vascular endothelial growth factor-A (VEGFA) by directly inhibiting TNF-α expression. Additionally, subsequent investigations identified that the ectopic level of VEGFA markedly abrogated the anti-angiogenic effect induced by miR-130b. Interestingly, VEGFA could in turn decrease the expression of miR-130b, thus forming a negative feedback loop that drives the angiogenesis of prostate cancer. CONCLUSION: These findings show that miR-130b/TNF-α/NF-κB/VEGFA feedback loop is significantly correlated with angiogenesis in prostate cancer and miR-130b could be regarded as potential therapeutic target for prostate cancer anti-angiogenesis treatment.
Subject(s)
Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , NF-kappa B/metabolism , Neovascularization, Pathologic/pathology , Prostatic Neoplasms/blood supply , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor A/metabolism , Animals , Apoptosis , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Cell Proliferation , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , NF-kappa B/genetics , Neovascularization, Pathologic/metabolism , Prognosis , Prostatic Neoplasms/genetics , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathology , Signal Transduction , Survival Rate , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/genetics , Vascular Endothelial Growth Factor A/genetics , Xenograft Model Antitumor AssaysABSTRACT
Frizzled 3 is an important receptor in the Wnt/ß-catenin pathway, a conserved signaling pathway that regulates gene expression and controls diverse developmental processes. However, the role of this protein during follicular development in the adult ovary is not known. The present study was designed to investigate the expression and localization of Frizzled 3 mRNA and protein during the estrous cycle in the mouse ovary through in situ hybridization (ISH), real-time quantitative polymerase chain reaction, immunohistochemistry and western blot. ISH results showed that in proestrus, high expression of Frizzled 3 was found in the granulosa and stroma with weak levels in the corpus luteum. In estrus and diestrus, the stroma had high Frizzled 3 expression, but levels were low in granulosa cells and corpus luteum. In the metestrus, moderate expression of Frizzled 3 was found in the stroma but low to no expression was found in luteal cells and follicles. The mRNA and protein levels of Frizzled 3 were found to be the highest in proestrus and diestrus compared to estrus and metestrus (P < 0.05), confirming the ISH results. During estrus and diestrus, high Frizzled 3 expression was observed in the stroma and moderate levels in granulosa cells, and during estrus and proestrus, low expression was seen in the oocyte cell membrane. The western blot results further confirmed this change during the estrous cycle. Together, these results indicate that Frizzled 3 is involved in regulating follicular development and oocyte maturation during the estrous cycle.
Subject(s)
Corpus Luteum/metabolism , Frizzled Receptors/genetics , Gene Expression Regulation, Developmental , Oocytes/metabolism , Ovarian Follicle/metabolism , Animals , Corpus Luteum/growth & development , Corpus Luteum/ultrastructure , Diestrus/genetics , Estrus/genetics , Female , Frizzled Receptors/metabolism , In Situ Hybridization , Mice , Mice, Inbred ICR , Oocytes/growth & development , Oocytes/ultrastructure , Ovarian Follicle/growth & development , Ovarian Follicle/ultrastructure , Proestrus/genetics , Real-Time Polymerase Chain Reaction , Time FactorsABSTRACT
The aim of this study was to investigate the role of cytokine genes in the susceptibility to Candida infection. A total of 275 consecutive patients diagnosed with Candida infection were selected between May 2010 and May 2011, along with 305 uninfected controls. Genotyping of the IL-1ß gene polymorphisms (IL1ß) rs1143634, IL1ßrs16944, IL8 rs4073, IL10 rs1800872, and IL10 rs1800896 was carried out using a 384-well plate format on the Sequenom MassARRAY platform. Patients with invasive Candida infections were more likely to have had an immunocompromised state, hematopoietic stem cell transplantation, solid organ transplant, solid tumor, chemotherapy within the past three months, neutropenia, surgery within the past 30 days, acute renal failure, liver failure, and/or median baseline serum creatinine. Conditional logistic regression analyses found that individuals with the rs1800896 GG genotype were associated with a higher risk of invasive Candida infections than those carrying the AA genotype (odds ratio = 0.61, 95% confidence interval = 0.37-0.94). From the results of this case-control study, we suggest that the cytokine IL-10 gene rs1800896 polymorphism might play a role in the etiology of invasive Candida infections.
Subject(s)
Candidiasis, Invasive/immunology , Genetic Predisposition to Disease , Immunocompromised Host , Interleukin-10/immunology , Interleukin-1beta/immunology , Interleukin-8/immunology , Polymorphism, Single Nucleotide , Acute Kidney Injury/genetics , Acute Kidney Injury/immunology , Acute Kidney Injury/microbiology , Acute Kidney Injury/surgery , Adult , Aged , Alleles , Candida/immunology , Candida/pathogenicity , Candidiasis, Invasive/genetics , Candidiasis, Invasive/microbiology , Candidiasis, Invasive/surgery , Case-Control Studies , Female , Gene Frequency , Humans , Interleukin-10/genetics , Interleukin-1beta/genetics , Interleukin-8/genetics , Liver Failure/genetics , Liver Failure/immunology , Liver Failure/microbiology , Liver Failure/surgery , Logistic Models , Male , Middle Aged , Neoplasms/genetics , Neoplasms/immunology , Neoplasms/microbiology , Neoplasms/surgeryABSTRACT
Mango is one of the most commercially important fruit crops in tropical and subtropical regions. To increase the efficiency of breeding strategies, two EST-derived marker systems were developed in the present study using information from the mango fruit transcriptome. Using simple sequence repeats, 218 of 230 primer pairs showed stable amplification for 7 mango genotypes with amplicons ranging from 84 to 160 bp; 93 of the primer pairs yielded polymorphic products. The proportion of polymorphic bands ranged from 16.67 to 100%, with a mean of 55.64%. In contrast, 86 primer pairs exhibited good amplification with clear bands for target region amplification polymorphism analysis, and a total of 66 primer combinations were polymorphic. These two novel sets of EST-derived markers will be of use in future studies of genetic diversity, genetic map construction, and marker-assisted selection in mango.