Abstract:
Objective: To investigate the influence of TGF-β and IFN-γ on the prol i feration, migration and invasion of melanoma cells. Methods:Melanoma cells were cultured in vitro. When tumor cells were confluent about 80% degree, cytokines were added into cel l cul ture media. The concentration of TGF-β and IFN-γ was 5ng/mL and 10ng/mL, respectively. Melanoma cel ls were divided into free-cytokine group, TGF-β group, IFN-γ group, TGF-β and IFN-γ group. Tumor cel ls in each group were then incubated for 8h, 16h, 24h, 32h, 40h and 48h, respectively. After incubation, fixing and staining with SRB, the optical densities and percentage viability were then determined by absorption at 540 nm (A 540 ). The scarification of tumor cells in each group on the surface was created by a 200 μ L pipette tube. The moti l i ty of tumor cel ls in each group was assessed by measuring the distance between scarifications. The speed of the scuffing closure was monitored after 12h. The invasive ability of melanoma cells was observed by transwell cultivation. The tumor cells that invaded through the Matrigel and adhered to the bottom of the outside membrane were determined by absorption at 595 nm (A 595 ). Gelatin zymography assay was used to examine the levels of matrix metalloproteinases- 2 (MMP- 2) and matrix metalloproteinases- 9 (MMP- 9) activity when the tumor cells were treated with cytokines after24h. MMP- 2 and MMP- 9 activity was demonstrated by gradation in the sodium dodecyl sulfate polyacryl -amide gel electrophoresis (SDS-PAGE) gelatin. MMP-2 and MMP- 9 activity was determined by Image analysis Software. Results: TGF-β promoted the prol i feration, migration and invasion of melanoma cel ls (P<0.05). However, IFN-γ inhibi ted the prol i feration, migration and invasion of melanoma cel ls (P<0.05). The effect weakened or disappeared when both of them were used (P>0.05). Conclusion:In vi tro, TGF-β may affect the inhibi tory effect of IFN-γ on the prol i feration, migration and invasion of melanoma cel ls. This study provided a better understanding of the relationship between tumor and inflammatory factors and established a good ba-sis for future research.