Table 1.
Correlation of IL-17A expression with clinicopathological features in 50 cervical cancer patients.
Figure 1.
IL-17A promoted cervical cancer cells migration and invasion.
(A, 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. (C) By cell invasive assay, the effect of IL-17A on cell invasion was detected (magnification 100×). (D) Total invasive cell number in each chamber was summarized. 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.
Figure 2.
IL-17A upregulated MMPs expression and activity and downregulated TIMP expression.
(A) MMPs expression was detected by real-time PCR analysis in the cervical cancer cells treated with and without IL-17A. (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. (C) Quantification of the protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. MMP2 (D) and MMP9 (E) concentrations in supernatants form cells treated with or without IL-17A were analyzed by ELISA. (F) The effects of IL-17A on the activities of MMP2 and MMP9 were analyzed by zymography assay. (G) Quantification of the activities of MMP-2 and MMP-9. 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.
Figure 3.
IL-17A can activate p38 and NF-κB signal pathways in cervical cancer cells.
Expression of p38 and p-p38 were detected by western blot analysis in C33A(A) and Caski(E) cells treated with or without IL-17A for 24 hours. Quantification of the protein levels of p38 and p-p38 in C33A(B) and Caski(F). 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(C) and Caski(G) cells treated with IL-17A (50 ng/mL) at indicated time points. Quantification of the nuclear protein levels of p50 and p65 in C33A(D) and Caski(H) cells. 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.
Figure 4.
Effects of p38 inhibitor (SB203580, SB), NF-κB inhibitor (PDTC), and IL-17A on cell invasion and MMP2, MMP9 expression in cervical cancer cells.
C33A(A) and Caski(E) 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) and Caski(F) cells was expressed as a percentage of control. C33A(C, D) and Caski(G, H) 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. * p<0.05 and ** p<0.01 compared with control group respectively.