Abstract:

[Abstract] Objective To explore the effects of zinc finger E-box binding protein (ZEB)23′UTR gene transfection
on proliferation, invasion and migration in human gastric epithelial cell line GES-1. Methods The synthetic ZEB23′UTR
and miR-200b micmics were transfected into GES-1cell line by lipofectamine2000. We set up control grop, the mutation group and ZEB23′UTR group. Real-time quantitative PCR was performed to evaluate the expression levels of miR-200a/b/c and ZEB1/ZEB2mRNAs after transfection．And then we set up control group, ZEB23′UTR group, ZEB23′UTR+negative control group and ZEB23′UTR+miR-200b micmics group. The protein expression levels of ZEB1, ZEB2, matrix metallopro?teinases (MMP) 2/9and proliferating cell nuclear antigen (PCNA) were detected by Western blot assay. The invasion and migration capability were analyzed by transwell assay and wound healing test. MTT assay was used to detect the proliferation ability.Results Compared with control group and mutation group, the expressions of miR-200a/b/c were significantly decreased, especially for miR-200b. And the expressions of ZEB1/ZEB2were significantly increased at both mRNA and protein levels after transfected with the ZEB23′ UTR, enhancing the capability of migration，invasion，and proliferation (P<0.05). Compared with ZEB23′UTR group, the capabilities of proliferation，invasion and migration were significantly lower in combined group.Conclusion ZEB23′ UTR can increase the ability of cell proliferation, invasion and metastasis through regulating the levels of miR-200a/b/c, and then influence the regulation of transcription of the target gene, which could lead to malignant transformation of GES-1cells.

Key words: genes, homeobox, 3′untranslated regions, microRNAs, gastric mucosa, epithelial cells, neoplasm metastasis, E盒结合锌指蛋白