基质金属蛋白酶-9反义寡核苷酸对卵巢癌细胞侵袭黏附能力的影响

Inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adherence of ovarian cancer cells

目的观察基质金属蛋白酶-9(MMP-9)反义寡核苷酸(ASODN)转染对卵巢癌细胞体外侵袭黏附行为的影响,并探讨其作用机制。方法以Lipofectinmin介导的MMP-9反义寡核苷酸转染至经纤黏连蛋白诱导MMP_9表达的卵巢癌细胞株HO-8910PM,利用RT．PCR、Western blot及明胶酶谱法检测转染寡核苷酸后HO-8910PM细胞MMP-9的mRNA、蛋白表达及酶活性的变化;通过细胞体外侵袭、迁移实验和黏附实验,检测细胞侵袭黏附能力的变化。结果卵巢癌细胞HO-8910PM转染MMP-9反义寡核苷酸后,MMP-9的mRNA及蛋白的表达受到抑制,抑制率分别为34．8％和42．5％,与对照组比较,差异有统计学意义(P<0．05);明胶酶活性也受到了抑制。反义寡核苷酸的转染降低了肿瘤细胞体外侵袭、迁移和黏附能力,侵袭和迁移抑制率分别为22．4％和24．8％,在60 min和90 min黏附抑制率分别为49．8％和38．3％。结论MMP-9反义寡核苷酸可抑制卵巢癌细胞的侵袭黏附能力,MMP-9有可能成为抗卵巢癌侵袭转移的分子靶点。

Objective To observe the inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adhesion ability in vitro of ovarian cancer cells, and to investigate the mechanisms of action. Methods MMP-9 antisense oligonucleotides were transfected by lipofectinmin into ovarian cancer cell line HO-8910PM cells expressing MMP-9 induced with fibronectin. RT-PCR, Western blot and gelatin zymography were used to detected MMP-9 expression of mRNA and protein and enzymatic activity. The ability of invasion and migration of ovarian cancer cells was assayed in Transwell cell culture chamber. Cell adhersion assay was carried out in a microculture well pre-coated with fibronectin. Results MMP-9 expressions of mRNA and protein were significantly decreased in the antisense-transfected cells. Comparing with the control group, the inhibition rate was 34.8% and 42.5%, respectively （ P 〈 0.05 ）. Its gelatin enzymatic activity was inhibited. Matrigel invasion assay and Transwell migration assay revealed markedly reduction in invasion and migration for the antisense group. The inhibition rates were 22.4% and 24.8%, respectively. The adhesion ability was also reduced. The inhibition rates were 49.8% and 38.3% at 60 min and 90 min, respectively. Conclusion MMP-9 down-regulation can significantly inhibit the ability of invasion and attachment of ovarian cells in vitro. MMP-9 may play an important role in invasion and metastasis of ovarian cells and potentially be a molecular target of blocking invasion and metastasis of ovarian cancer.