Hu Xiao-xia, Li Li, Li Dan-rong, Zhang Wei, Tang Bu-jian
Department of Gynecological Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China.
Zhonghua Zhong Liu Za Zhi. 2006 Sep;28(9):662-5.
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.
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.
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.
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.
观察反义MMP - 9寡脱氧核苷酸对卵巢癌细胞体外侵袭和黏附能力的抑制作用,并探讨其作用机制。
采用脂质体介导将MMP - 9反义寡核苷酸转染至经纤连蛋白诱导表达MMP - 9的卵巢癌细胞系HO - 8910PM细胞。运用RT - PCR、Western blot和明胶酶谱法检测MMP - 9 mRNA和蛋白表达及酶活性。在Transwell细胞培养小室中检测卵巢癌细胞的侵袭和迁移能力。在预先包被纤连蛋白的微孔培养板中进行细胞黏附实验。
反义转染细胞中MMP - 9的mRNA和蛋白表达显著降低。与对照组相比,抑制率分别为34.8%和42.5%(P <0.05)。其明胶酶活性受到抑制。基质胶侵袭实验和Transwell迁移实验显示反义组的侵袭和迁移明显减少。抑制率分别为22.4%和24.8%。黏附能力也降低。在60分钟和90分钟时的抑制率分别为49.8%和38.3%。
下调MMP - 9可显著抑制卵巢癌细胞体外侵袭和黏附能力。MMP - 9可能在卵巢癌细胞侵袭和转移中起重要作用,有望成为阻断卵巢癌侵袭和转移的分子靶点。