Influence Of Interleukin-23on Angiogenesis In Mouse Malignant Melanoma

Chronic inflammation has long been associated with increased incidence of malignancy and similarities in the regulatory mechanisms have been suggested for more than a century. Infiltration of innate immune cells, elevated activities of matrix metalloproteases and increased angiogenesis and vasculature density are a few examples of the similarities between chronic and tumour-associated inflammation2. Conversely, the elimination of early malignant lesions by immune surveillance, which relies on the cytotoxic activity of tumour-infiltrating T cells or intra-epithelial lymphocytes, is thought to be rate-limiting for the risk to develop cancer3. Here we show a molecular connection between the rise in tumour-associated inflammation and a lack of tumour immune surveillance.Local tumor growth factor and cytokine expression not only help protect tumor tissue remodeling and angiogenesis, but also resist to from the immune system. Accordingly, anti-tumor and tumor-promoting role of chronic inflammation, is closely related to release of inflammation cytokines. This study established early in the laboratory animal model of chronic inflammatory melanoma composite based on the detection of mouse serum cytokine content. Combining the results obtained in the previous animal model, we analyse the correlation of interleukin (IL)-6, IL-17, IL-23, vascular endothelial growth factor (VEGF) and tumor microvessel density (MVD).We select inflammatory factor IL-23in vitro experiments to observe the B16F10cell proliferation, migration, invasion and pipe forming ability of (mimicry vascular), The main contents are as follows:1. Detection of cytokines in the serum of mice and screeningThis part of the study, select mouse serum (taken from the laboratory pre-established chronic inflammation-melanoma the compound animal models) as a detection target, Some cytokines are detected by enzyme-linked immunosorbent assay (ELISA) and analysed by health statistical methods.The results are as follows:Four kinds (IL-6,IL-17,IL-23,VEGF) of cytokines contained in mouse serum of I+T group are greater than T group except IL-17and IL-23on the9th day(P<0.05or P<0.01). Also these cytokines have a kind of positive correlation with micro-vessels density. The levels of IL-6and IL-23are positively correlated with VEGF.According to these results and the current research condition between IL-23and malignant melanoma, we choose IL-23as object to do further studies.2. The influence of IL-23on proliferation, migration, invasion and pipe forming ability of B16F10cells in vitro.In order to observe the influence of inflammatory cytokines on proliferation, migration, invasion and pipe forming ability of tumor cells.In this part, we choose IL-23and mouse-derived B16F10cell as our study object. We add exogenous IL-23in the medium of B16F10, to observe its influence on proliferation, migration, invasion and pipe forming ability.The result of MTT test tells us:the proliferation of tumor cells is higher than control group under the intervention of IL-23and the difference has a statistical significant(P<0.05). Also the shape of B16F10cells is different from the intervened group.The migration distance of the mouse melanoma B16F10cells was significantly longer than that of control group by wound-healing assay(P<0.05), when treated with various concentrations of IL-23.The B16F10cells treated with various concentrations of IL-23were found through the membrane more than the control group in a Transwell assay(P<0.05).The activity of MMP-2and MMP-9of the mouse melanoma B16F10cells was increased after treated with IL-23by Gelatin zymography assay(P<0.05).The ability of pipe formation of the mouse melanoma B16F10cells was increased significantly in three-dimensional cultures when treated with various concentrations of IL-23.The expression of NF-κB of the mouse melanoma B16F10cells were increased significantly by Western blot when treated with IL-23(P<0.05).In summary, we detect the volume of cytokines in mouse serum by enzyme-linked immunosorbent assay (ELISA) and analyse the correlation between cytokins and micro-vessels by statistical methods. We choose IL-23as our study object in vitro, we test the influence of IL-23on proliferation, migration, invation and pipe forming ability of B16F10cells. Our conclusions are as follows: 1. Some kinds of cytokines released from chronic inflammation area have a close correlation in tumor angiogenesis. Four kinds (IL-6,IL-17,IL-23,VEGF) of cytokines contained in mouse serum have a kind of positive correlation with micro-vessels density. The levels of IL-6and IL-23are positively correlated with VEGF.2.The impact of IL-23on B16F10cells:according to the results of MTT test, Transwell test, Gelatin zymography assay and three-dimensional cultures, we know that IL-23can promote the proliferation, migration and invasion ability, and IL-23can induced the pipe forming ability of B16F10cells.