Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P10145 (IL-8)
 23,849 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The human interleukin-17F(hIL-17F) gene was amplified by RT-PCR from PHA-activated human peripheral blood mononuclear cells (PBMCs). It was then subcloned into the retrovirus vector pSIV-1. The pSIV-1/hIL-17F together with its two-helper virus vectors pHIT456 and pHIT60 cotransfected into the package cell 293T by lipofectin to produce mature recombinant retrovirus, which was then used to infect SMMC-7721 hepatocarcinoma cells (HCCs), and the cells were selected in the presence of G418. The integration, transcription, expression of hIL-17F gene in SMMC-7721 cells was identified by PCR, RT-PCR and Western blot respectively. MTT and FCM showed that hIL-17F couldn't alter the proliferation and cell cycle of SMMC-7721 cells, but ELISA showed that it could down-regulate IL-6, IL-8 and VEGF expression. The effect of rhIL-17F supernatant on growth suppressing of ECV304 cells was observed by MTT. The experiment of human hepatocarcinoma xenograft tumor in nude mice showed that the formation and growth rates of hIL-17F-transgenic SMMC-7721 showed an obvious decline, and VEGF and CD34 expression and angiogenesis of the transgenic neoplasms was also evidently defined. hIL-17F can markedly inhibit the growth of human hepatocarcinoma xenograft tumor in nude mice by antiangiogenesis. This study provided an experimental evidence for further conducting tumor gene therapy by targeting vascularity and exploiting antiangiogenic novel medicine related to hIL-17F.
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PMID:[The effect of human IL-17F on growth of human hepatocarcinoma xenograft tumor in nude mice]. 1703

Carbon nanotube-based nanovectors, especially functionalized nanotubes, have shown potential for therapeutic drug delivery. 6-Aminohexanoic acid-derivatized single-wall carbon nanotubes (AHA-SWNTs) are soluble in aqueous stock solutions over a wide range of physiologically relevant conditions; however, their interactions with cells and their biological compatibility has not been explored. Human epidermal keratinocytes (HEKs) were dosed with AHA-SWNTs ranging in concentration from 0.00000005 to 0.05 mg/ml. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability decreased significantly (p < .05) from 0.00005 to 0.05 mg/ml after 24 h. The proinflammatory mediators of inflammation cytokines interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-alpha, IL-10, and IL-1beta were also assessed. Cytokine analysis did not show a significant increase in IL-6 and IL-8 in the medium containing 0.000005 mg/ml of AHA-SWNTs from 1 to 48 h. IL-6 increased in cells treated with 0.05 mg/ml of AHA-SWNTs from 1 to 48 h, whereas IL-8 showed a significant increase at 24 and 48 h. No significant difference (p < .05) was noted with TNF-alpha, IL-10, and IL-1beta expression at any time point. Transmission electron microscopy of HEKs treated with 0.05 mg/ml AHA-SWNTs for 24 h depicted AHA-SWNTs localized within intracytoplasmic vacuoles in HEKs. Treatment with the surfactant 1% Pluronic F127 caused dispersion of the AHA-SWNT aggregates in the culture medium and less toxicity. These data showed that the lower concentration of 0.000005 mg/ml of AHA-SWNTs maintains cell viability and induces a mild cytotoxicity, but 0.05 mg/ml of AHA-SWNTs demonstrated an irritation response by the increase in IL-8.
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PMID:Biological interactions of functionalized single-wall carbon nanotubes in human epidermal keratinocytes. 1745 50

The placenta is an attractive new source of mesenchymal stem cells (MSCs), but the biological characteristics of placenta-derived MSCs (P-MSCs) have not yet been characterized. We successfully isolated, cultured and expanded P-MSCs using routine methods. Under appropriate induction conditions, these cells can differentiate into bone, cartilage, fat and hepatocyte-like cells. In addition, the proliferative response of P-MSCs to different cytokines was monitored using the MTT assay. The results show that low concentrations of proinflammatory cytokines, e.g. RANTES, interleukin (IL)-1, IL-6 and IL-8 can stimulate the proliferation of P-MSCs in a dose-dependent manner, peaking at concentrations of 40 ng/ml of RANTES, 10 ng/ml of IL-1 and IL-6, and 150 ng/ml of IL-8 (p < 0.01). The level of proliferation decreased when the concentration of these four cytokines increased beyond these values. On the other hand, anti-inflammatory cytokines hepatocyte growth factor and IL-4 had an inhibitory effect on P-MSCs. In conclusion, the placenta contains MSCs that are consistent with the characteristics of bone marrow MSCs. Low concentrations of proinflammatory chemokines stimulated the proliferation of P-MSCs while anti-inflammatory cytokines inhibited the growth of P-MSCs in a dose-dependent manner.
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PMID:Biological characteristics of human placental mesenchymal stem cells and their proliferative response to various cytokines. 1763 Apr 77

Embryonic development associated gene (EDAG), which is overexpressed in hematopoietic neoplasms and leukemia cell lines, has been reported to participate in the leukemia cell differentiation and proliferation. This study investigated whether retrovirus-mediated transfer of a siRNA against EDAG can reduce the growth of leukemia cells which highly express EDAG in vitro and in vivo. The stable transfected cells were identified with RT-PCR, the effect of EDAG/siRNA on the growth of the human erythroleukemia cell line HEL was analyzed by MTT assay, and angiogenic factor IL-8 release was evaluated by ELISA and RT-PCR. The results showed that EDAG/siRNA can silence the expression of EDAG in HEL cells. Down-regulation of EDAG expression by retrovirus-mediated siRNA inhibited the cell proliferation and tumor growth. Knockdown of EDAG expression by siRNA is also associated with decreased expression of the anti-angiogenic factor IL-8, suggesting that EDAG stimulates tumor growth at least in part by regulating angiogenesis. This study suggests that siRNA-mediated gene silencing of EDAG could potentially be a therapeutic strategy for EDAG over-expressing leukemia cells.
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PMID:Down-regulation of EDAG expression by retrovirus-mediated small interfering RNA inhibits the growth and IL-8 production of leukemia cells. 1767 16

To explore effects of Forsythia koreana methanol extract (FKME) on mast cell-mediated allergic and inflammatory properties, the effect of FKME was evaluated on compound 48/80-induced systemic anaphylaxis, ear swelling, and anti-dinitrophenyl (DNP) immunoglobulin E (IgE)-induced passive cutaneous anaphylaxis (PCA). In addition, the effect of FKME was investigated on the histamine release from rat peritoneal mast cells (RPMCs) stimulated by compound 48/80, which promotes histamine release. The human mast cell line HMC-1 was stimulated by phorbol 12-myristate 13-acetate plus calcium ionophore A23187. Activated HMC-1 can produce several proinflammatory and chemotactic cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, and IL-8. Cytokine levels in the culture supernatant were measured by an enzyme-linked immunosorbent assay. Cytotoxicity by FKME was determined by a 3-(4,5-dimethylthiazol-2-yl)-diphenyl-tetrazolium bromide (MTT) assay. FKME inhibited compound 48/80-induced systemic anaphylactic shock and ear swelling in mice. When 1 g/kg FKME was pretreated or posttreated with mice, compound 48/80-induced mice morality was 50 and 66.7%, respectively. One gram per kilogram of FKME pretreatment inhibited ear-swelling responses derived from compound 48/80 by 29.75%. A PCA reaction was inhibited by 17.9%. In an in vitro model, FKME (1 mg/ml) inhibited histamine release from the RPMCs by 13.8% and TNF-alpha, IL-6, and IL-8 production from HMC-1 cells by 71.16% (P < 0.001), 86.72% (P < 0.001), and 44.6%, respectively. However, FKME had no cytotoxic effects on cell viability. In conclusion, FKME inhibited not only systemic anaphylaxis and ear swelling induced by compound 48/80 but also inhibited a PCA reaction induced by anti-DNP IgE in vivo. Treatment with FKME showed significant inhibitory effects on histamine, TNF-alpha, IL-6, and IL-8 release from mast cells.
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PMID:Observations of Forsythia koreana methanol extract on mast cell-mediated allergic reactions in experimental models. 1772 24

Previous work from our laboratory has demonstrated overexpression of chemokines in head and neck cancer, and the utility of targeting CXCL5 for tumor therapy in a preclinical model. In the present study, we investigated the contribution of a related chemokine, CXCL8, to cellular properties associated with tumor progression, namely cell growth and motility. Expression of CXCL8 was detectable in multiple squamous carcinoma cell lines, indicating a possible role in pathogenesis. Overexpression of CXCL8 in HN4 primary tumor cells with low endogenous CXCL8 levels was found to increase cell growth, as judged by cell counting and MTT assays. Conversely, RNAi-mediated knockdown of CXCL8 expression in HN12 cells, derived from a synchronous metastasis and which express high levels of this chemokine, resulted in a decrease in proliferation. Similarly, overexpression of CXCL8 enhanced migration of HN4 cells, while suppression of CXCL8 inhibited HN12 cell migration and invasion through a basement membrane substitute. Taken together, these findings support the hypothesis that CXCL8 affects multiple processes involved in tumor progression and identify CXCL8 as a potential therapeutic target, similar to CXCL5.
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PMID:Roles of CXCL8 in squamous cell carcinoma proliferation and migration. 1828 85

Testing chemicals for their ability to cause skin irritation is required for all ingredients of products that come into contact with the skin. Here, we describe a potential method for determining the irritant potency of a chemical in vitro and apply the method to two different reconstructed epidermis models which exhibit different barrier properties. Two surfactants: sodium dodecyl sulphate, Triton X100 and two non-surfactants: 2-4-di-nitro-chloro-benzene, cinnamaldehyde were applied topically in a dose response for 24h. Biomarkers IL-1alpha, IL-1RA, IL-8 and MTT were assessed and EC(50) values determined. Variation in barrier properties between the epidermal models led to variation in the extent of penetration of surfactants, but not of non-surfactants which in turn influenced the EC(50) value obtained from surfactants. Furthermore, EC(50) values showed that no single biomarker could be classed as the most sensitive biomarker since biomarker sensitivity differed between the different chemicals studied. However, the ranking of the chemicals in order of strong to weak irritant was the same irrespective of the model used and also independent of the biomarker used (Triton X100>DNCB>SDS>CA). This study describes a method which not only distinguishes an irritant from a non-irritant but which may possibly also be used to determine irritant potency.
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PMID:Potential method to determine irritant potency in vitro - Comparison of two reconstructed epidermal culture models with different barrier competency. 1913 41

Inflammation is a major contributing factor to many blinding disorders including uveitis, diabetic retinopathy, and age-related macular degeneration. Here we examined the response of the retinal pigment epithelium (RPE) to physiological levels of lipopolysaccharide (LPS) to understand the role of this epithelium in inflammatory retinal conditions. Expression of a group of inflammatory mediators was identified by gene array analysis and confirmed by PCR and immunocytochemistry in primary human RPE cultures and ARPE19. The effects of LPS on the expression of these cytokines and RPE survival were examined by PCR, Luminex bead, and MTT assays. RPE cells express many cytokine receptors including IL-1R, -4R, -6R, -8RA, IFNAR1, IFNGR1/2 and secrete a range of pro- and anti-inflammatory cytokines including IL-4, -6, -8, -10, -17, IFN-gamma, MCP-1, and VEGF. LPS increases IL-13RA1 and IFNAR1, and decreases IL-7R receptor expression. It also increases RPE secretion of IL-4, -6, -8, -10, IFN-gamma and MCP-1, and is toxic to RPE cells at LC(50)=17.7 microg/ml. LPS toxicity is mediated by IL-6 and IL-8 through an autocrine feedback loop. Silencing IL-6R and IL-8RA gene expression by siRNA blocks death by their respective ligands or LPS. These findings imply that RPE cells are acutely sensitive to inflammatory stress and that over secretion of IL-6 and IL-8 by this epithelium during inflammatory stimulus may be an underlying factor in the progression of some retinal pathologies.
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PMID:Bacterial endotoxin activates retinal pigment epithelial cells and induces their degeneration through IL-6 and IL-8 autocrine signaling. 1915 52

Irritant contact dermatitis is the result of an innate inflammatory response of the skin to direct injury. It is caused by a single, repeated or continued application of an irritant, with the source most often being a chemical. Therefore, European regulations require strict screening of all ingredients in consumer products. Until recently, identifying a potential irritant has completely relied on animal testing (for example, Draize test). Besides the ethical problems, both the 7th Amendment to the Cosmetics Directive and Registration, Evaluation and Authorization of Chemicals legislation have stimulated the development of alternative tests for the assessment of potential toxicological effects of substances. This review is aimed at describing current in vitro skin irritation models and the biomarkers used to assess the degree of irritancy of a potential irritant. Four models are described: keratinocyte and fibroblast cultures grown under submerged culture conditions, epidermal equivalents, skin equivalents and freshly isolated skin. Biomarkers such as IL-1alpha, IL-6, IL-8, PGE2, SKALP, HSP70 and kinases are described along with changes in metabolic activity (MTT assay) and cytosolic leakage (lactate dehydrogenase assay). Noticeable is the limited number of genomic and proteomic studies. Such studies have the potential to identify novel biomarkers and to elucidate the mechanism of irritant contact dermatitis.
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PMID:In vitro irritation models and immune reactions. 1918 58

Inflammation is a complex process involving cytokine production to regulate host defense cascades. In contrast to the therapeutic significance of acute inflammation, a pathogenic impact of chronic inflammation on cancer development has been proposed. Upregulation of inflammatory cytokines, such as IL-1beta and IL-8, has been noted in prostate cancer patients and IL-8 has been shown to promote prostate cancer cell proliferation and migration; however, it is not clear whether IL-1beta regulates IL-8 expression in prostate cancer cells. Glucosamine is widely regarded as an anti-inflammatory agent and thus we hypothesized that if IL-1beta activated IL-8 production in prostate cancer cells, then glucosamine ought to blunt such an effect. Three prostate cancer cell lines, DU-145, PC-3, and LNCaP, were used to evaluate the effects of IL-1beta and glucosamine on IL-8 expression using ELISA and RT-PCR analyses. IL-1beta elevated IL-8 mRNA expression and subsequent IL-8 secretion. Glucosamine significantly inhibited IL-1beta-induced IL-8 secretion. IL-8 appeared to induce LNCaP cell proliferation by MTT assay; involvement of IL-8 in IL-1beta-dependent PC-3 cell migration was demonstrated by wound-healing and transwell migration assays. Inhibitors of MAPKs and NFkappaB were used to pinpoint MAPKs but not NFkappaB being involved in IL-1beta-mediated IL-8 production. IL-1beta-provoked phosphorylation of all MAPKs was notably suppressed by glucosamine. We suggest that IL-1beta can activate the MAPK pathways resulting in an induction of IL-8 production, which promotes prostate cancer cell proliferation and migration. In this context, glucosamine appears to inhibit IL-1beta-mediated activation of MAPKs and therefore reduces IL-8 production; this, in turn, attenuates cell proliferation/migration.
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PMID:Glucosamine inhibits IL-1beta-mediated IL-8 production in prostate cancer cells by MAPK attenuation. 1962 64


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