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目的探讨内皮抑制素对体外培养的黑素瘤A375细胞核因子κB(NF-κB)和基质金属蛋白酶-9(MMP9)表达的影响。方法 CCK-8法检测内皮抑制素对A375细胞增殖的抑制作用;Transwell小室试验检测内皮抑制素对A375细胞侵袭能力的影响;逆转录聚合酶链反应(RT-PCR)和Western-blot法分别检测内皮抑制素对A375细胞NF-κB,MMP9 mRNA和二者蛋白表达的影响。结果内皮抑制素对A375细胞增殖抑制作用呈时间-浓度依赖性;20μg/mL内皮抑制素作用A375细胞72h后:细胞穿膜数(21.5±7.3)低于对照组(56.8±6.2);胞内NF-κB,MMP9 mRNA以及二者蛋白表达水平均有明显下降,与对照组相比的差异均有统计学意义(P均<0.05)。结论内皮抑制素可下调NF-κB和MMP9的表达,从而抑制黑素瘤浸润和转移。这为内皮抑制素临床治疗皮肤黑素瘤提供了部分理论依据。
Objective To investigate the effect of endostatin on the expression of nuclear factor κB (NF-κB) and matrix metalloproteinase-9 (MMP9) in cultured melanoma A375 cells. Methods The inhibitory effect of endostatin on the proliferation of A375 cells was detected by CCK-8 assay. The effect of endostatin on the invasiveness of A375 cells was detected by Transwell chamber assay. The expression of endostatin was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blot respectively Effects of endostatin on the expressions of NF-κB, MMP9 mRNA and their proteins in A375 cells. Results Endostatin inhibited the proliferation of A375 cells in a time-and-concentration-dependent manner. After treated with 20 μg / mL of endostatin for 72 h, the number of cells penetrating through the membrane was significantly lower (21.5 ± 7.3) than that of the control (56.8 ± 6.2) The expression of NF-κB, MMP9 mRNA and their protein levels both decreased significantly compared with the control group (all P <0.05). Conclusion Endostatin can down-regulate the expression of NF-κB and MMP9, thereby inhibiting melanoma invasion and metastasis. This provides a partial theoretical basis for the clinical treatment of cutaneous melanoma with endostatin.