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探讨O-GlcNAc糖基转移酶(OGT)介导的O-GlcNAc糖基化在卵巢癌细胞迁移过程中的作用及其分子机制。采用RNAi基因干扰技术,干扰OGT基因的表达,构建低表达O-GlcNAc糖基化的卵巢癌HO-8910PM细胞模型;采用O-GlcNAc糖苷酶(OGA)抑制剂PUGNAc或Thiamet G诱导,构建高表达O-GlcNAc糖基化的卵巢癌OVCAR3细胞模型;通过qPCR和Western blot法验证细胞模型的有效性;通过体外细胞迁移实验观察O-GlcNAc糖基化对卵巢癌细胞迁移的影响;并通过qPCR法进一步检测不同的细胞模型中基质金属蛋白酶-2(MMP-2)和MMP-9的表达。结果显示,在HO-8910PM细胞中,干扰OGT基因的表达,可以降低MMP-2和MMP-9的mRNA水平,抑制HO-8910PM细胞的迁移。综上,OGT介导的O-GlcNAc糖基化可以通过调节MMP-2和MMP-9的表达参与调控卵巢癌细胞的迁移。
To investigate the role of O-GlcNAc glycosyltransferase (OGT) -mediated O-GlcNAc glycosylation in ovarian cancer cell migration and its molecular mechanism. Overexpression of OGT gene was performed by interfering with the expression of OGT gene and RNAi knockdown was used to construct ovarian cancer HO-8910PM cell model with low expression of O-GlcNAc. Overexpression of OGlcNAc or Thiamet G was induced by overexpression of OGlcNAc, O-GlcNAc-overexpressing ovarian cancer OVCAR3 cell model; validating the cell model by qPCR and Western blot; observing the effect of O-GlcNAc glycosylation on ovarian cancer cell migration by in vitro cell migration assay; Further testing for the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 in different cell models. The results showed that knocking down the expression of OGT gene in HO-8910PM cells could decrease the mRNA level of MMP-2 and MMP-9 and inhibit the migration of HO-8910PM cells. In summary, OGT-mediated O-GlcNAc glycosylation can regulate the migration of ovarian cancer cells by regulating the expression of MMP-2 and MMP-9.