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目的探讨血管内皮生长因子(VEGF)对人黑素瘤细胞SK-MEL-1侵袭力以及对该细胞中基质金属蛋白酶9(MMP-9)的影响,初步研究VEGF对肿瘤侵袭能力的影响及可能的作用机制。方法使用VEGF体外培养人黑素瘤SK-MEL-1细胞,免疫组化法检测该细胞VEGFR-1水平;通过细胞体外侵袭实验检测细胞侵袭能力的改变,再分别使用含20,40,80μg/LVEGF培养液培养SK-MEL-1细胞,以正常培养SK-MEL-1细胞为空白对照组,使用半定量RT-PCR和Western blot法对4组细胞中MMP-9 mRNA表达和蛋白水平进行分析。结果 VEGFR-1在SKM-EL-1细胞胞核和胞浆中均有表达。外加VEGF培养后,细胞侵袭力明显增强(P<0.01);MMP-9mRNA表达和蛋白水平在外加VEGF组明显高于空白对照组,且高浓度和低浓度组之间差异有统计学意义(P均<0.05)。结论人黑素瘤SK-MEL-1细胞系中存在有自分泌机制,VEGF可通过自分泌机制上调人黑素瘤SK-MEL-1细胞MMP-9mRNA表达及蛋白水平,进而促进肿瘤的侵袭转移。
Objective To investigate the effect of vascular endothelial growth factor (VEGF) on the invasiveness of human melanoma SK-MEL-1 cells and the effect of VEGF on tumor invasiveness and MMP-9 The mechanism of action. Methods The human melanoma SK-MEL-1 cells were cultured with VEGF in vitro and the level of VEGFR-1 was detected by immunohistochemistry. The invasiveness of cells was evaluated by in vitro invasion assay. The cells were treated with 20, 40, 80μg / The SK-MEL-1 cells were cultured in LVEGF culture medium, and the SK-MEL-1 cells were cultured as blank control group. The expression of MMP-9 mRNA and protein in four groups of cells were analyzed by semi-quantitative RT-PCR and Western blot . Results VEGFR-1 was expressed in the nuclei and cytoplasm of SKM-EL-1 cells. Compared with blank control group, the expression of MMP-9 mRNA and protein in the VEGF-treated group was significantly higher than that in the blank control group (P <0.01), and the difference between the high and low concentration groups was statistically significant (P <0.01) All <0.05). Conclusion Autocrine mechanism exists in human melanoma SK-MEL-1 cell line. VEGF can up-regulate the expression of MMP-9 mRNA and protein in melanoma SK-MEL-1 cells by autocrine mechanism, .