IL-17A promotes gastric cancer cell migration and invasion.

<p>(A) IL-17A treated GC cells (AGS, BGC-823 and SGC-7901) showed higher motility in a wound-healing assay, compared with cells without IL-17A treatment. (B) The percent migration rate is expressed as a percentage of the beginning area. (C) Effect of IL-17A on cell invasion was detected by transwell assay. Representative pictures of cells migrated through Matrigel-coated transwell were shown. (D) Total invasive cell number in each chamber was summarized as a percentage of control. Values represent the means ± SD of three independent experiments performed in triplicate. *<i>p</i><0.05 and **<i>p</i><0.01 compared with the control group.</p

IL-17A activates NF-κB pathway in AGS cells.

<p>(A) Western blotting analysis was used to detect overall p50, p65, p52, c-Rel and RelB expression in AGS cells treated with IL-17A (50 ng/ml) at indicated time points. (B) Quantification of the protein levels of overall p50, p65, p52, c-Rel and RelB. (C) Western blotting analysis was used to detect nuclear p50, p65, p52, c-Rel and RelB expression in AGS cells treated with IL-17A (50 ng/mL) at indicated time points. (D) Quantification of the protein levels of nuclear p50, p65, p52, c-Rel and RelB. (E) The relative ratio of nuclear to overall fraction of p50, p65, p52, c-Rel and RelB. Values represent the means ± SD of three independent experiments performed in triplicate. **<i>p</i><0.01, compared with the control group.</p

Interleukin 17A Promotes Gastric Cancer Invasiveness via NF-κB Mediated Matrix Metalloproteinases 2 and 9 Expression

<div><p>Interleukin 17A (IL-17A), as a pro-inflammatory cytokine, is involved in pathology of inflammatory diseases and tumor microenvironment. The aim of this study is to investigate the effect of IL-17A on the invasiveness of gastric cancer (GC). In the study, we found that IL-17A could promote the migration and invasion of GC cells. Furthermore, after treated with IL-17A, the expressions and activities of matrix metalloproteinase 2 (MMP-2) and MMP-9 were upregulated, while the expressions of TIMP-1 and TIMP-2 were downregulated. Moreover, the nuclear/overall fractions of p65 and p50 were dramatically elevated by IL-17A. Pretreatment with helenalin, a nuclear factor-κB (NF-κB) inhibitor, was proved to abolish the promoting effect of IL-17A on the invasion ability of GC cells and upregulation of MMP-2 and MMP-9. In conclusion, our findings illustrated that IL-17A could promote the invasion of GC cells by activating NF-κB pathway, and subsequently upregulating the expression of MMP-2 and MMP-9. These results may lead to the identification of new diagnostic markers and therapeutic targets of GC.</p></div

IL-17A promotes the expressions and activities of MMP-2 and MMP-9 and suppresses the expressions of TIMP-1 and TIMP-2 in gastric cancer cells.

<p>(A) Expressions of MMPs in GC cells (AGS and BGC-823) were compared by western blotting between cells treated with and without IL-17A (50 ng/ml) for 24 h. (B) Quantification of the protein levels of MMP-2 and MMP-9. (C) Effects of IL-17A on the activities of MMP-2 and MMP-9. (D) Quantification of the activities of MMP-2 and MMP-9. Values represent the means ± SD of three independent experiments performed in triplicate. *<i>p</i><0.05 and **<i>p</i><0.01 compared with the control group.</p

Effects of the NF-κB inhibitor and IL-17A on cell invasion and the expressions of MMP-2 and MMP-9 in AGS cells.

<p>(A) 1×10⁶ AGS cells were pretreated with helenalin (5 µM) for 30 min and then incubated in the presence or absence of IL-17A (50 ng/ml) for 24 h. Cellular invasiveness was measured using the transwell invasion assay. (B) The percent invasion rate was expressed as a percentage of control. (C, D) AGS cells were treated and then subjected to western blotting to analyze the protein levels of MMP-2 and MMP-9. Values represent the means ± SD of three independent experiments performed in triplicate. *<i>p</i><0.05 and **<i>p</i><0.01 compared with the control group.</p

IL-17A can activate p38 and NF-κB signal pathways in cervical cancer cells.

<p>Expression of p38 and p-p38 were detected by western blot analysis in C33A(<b>A</b>) and Caski(<b>E</b>) cells treated with or without IL-17A for 24 hours. Quantification of the protein levels of p38 and p-p38 in C33A(<b>B</b>) and Caski(<b>F</b>). Values are represented as means ± SD of three independent experiments performed in triplicate. *p<0.05 and **p<0.01 compared with control group, respectively. Western blot analysis was used to detect nuclear p50 and p65 expression in C33A(<b>C</b>) and Caski(<b>G</b>) cells treated with IL-17A (50 ng/mL) at indicated time points. Quantification of the nuclear protein levels of p50 and p65 in C33A(<b>D</b>) and Caski(<b>H</b>) cells. Values are represented as means ± SD of three independent experiments performed in triplicate. * <i>p</i><0.05 and ** <i>p</i><0.01 compared with control group respectively.</p

Correlation of IL-17A expression with clinicopathological features in 50 cervical cancer patients.

<p>*Partial data unavailable, statistics was done on the available data.</p><p>Correlation of IL-17A expression with clinicopathological features in 50 cervical cancer patients.</p

Effects of p38 inhibitor (SB203580, SB), NF-κB inhibitor (PDTC), and IL-17A on cell invasion and MMP2, MMP9 expression in cervical cancer cells.

<p>C33A(<b>A</b>) and Caski(<b>E</b>) cells were pretreated with SB (20 µM) and PDTC for 30 min, then incubated in the presence or absence of IL-17A (50 ng/mL) for 24 h. The cell invasive abilities were performed by Boyden chamber invasion assay. The percentage of invasive rate of C33A(<b>B</b>) and Caski(<b>F</b>) cells was expressed as a percentage of control. C33A(<b>C, D</b>) and Caski(<b>G, H</b>) cells were treated and then subjected to western blot to analyze the protein levels of MMP2, MMP 9. Values are represented as means ± SD of three independent experiments performed in triplicate. * <i>p</i><0.05 and ** <i>p</i><0.01 compared with control group respectively.</p

IL-17A promoted cervical cancer cells migration and invasion.

<p>(<b>A, B</b>) Compared with control group cells, IL-17A treated cervical cancer cells (HeLa, C33 A, and Caski) showed higher motility in a wound-healing assay. (<b>C</b>) By cell invasive assay, the effect of IL-17A on cell invasion was detected (magnification 100×). (<b>D</b>) Total invasive cell number in each chamber was summarized. Values are represented as means ± SD of three independent experiments performed in triplicate. * <i>p</i><0.05 and ** <i>p</i><0.01 compared with control group respectively.</p

IL-17A upregulated MMPs expression and activity and downregulated TIMP expression.

<p>(<b>A</b>) MMPs expression was detected by real-time PCR analysis in the cervical cancer cells treated with and without IL-17A. (<b>B</b>) After treating with IL-17A for 24 hours, the expression of MMP2, MMP9, TIMP-1, and TIMP-2 were detected by western blot analysis in cervical cancer cells. (<b>C</b>) Quantification of the protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. MMP2 (<b>D</b>) and MMP9 (<b>E</b>) concentrations in supernatants form cells treated with or without IL-17A were analyzed by ELISA. (<b>F</b>) The effects of IL-17A on the activities of MMP2 and MMP9 were analyzed by zymography assay. (<b>G</b>) Quantification of the activities of MMP-2 and MMP-9. Values are represented as means ± SD of three independent experiments performed in triplicate. * <i>p</i><0.05 and ** <i>p</i><0.01 compared with control group respectively.</p