CAJ | 학술논문

目的：探讨E盒结合锌指蛋白（ZEB）2基因3′非翻译区（3′UTR）转染人胃黏膜上皮细胞GES-1后对其增殖、侵袭、迁移的影响。方法将人工合成的ZEB23′UTR质粒及miR-200b micmics采用脂质体Lipofectamine 2000转染GES-1细胞。分别设对照组、突变组和ZEB23′UTR组，qRT-PCR检测转染后miR-200a/b/c及ZEB1/ZEB2 mRNA的表达。再设对照组、ZEB23′UTR组、ZEB23′UTR+无义序列组和ZEB23′UTR+miR-200b micmics组。West-ern blot检测转染后ZEB1/ZEB2、基质金属蛋白酶（MMP）-2/9、增殖细胞核抗原（PCNA）蛋白的表达；Transwell侵袭实验和划痕实验检测细胞侵袭迁移能力；MTT法检测细胞增殖活性。结果与对照组及突变组相比，转染ZEB23′UTR组miR-200a/b/c的表达均下调，以miR-200b最为明显，ZEB1/ZEB2 mRNA及蛋白水平表达上调，其细胞迁移、侵袭、增殖活性均增强（P<0.05）。ZEB23′UTR+miR-200b micmics组较ZEB23′UTR组迁移、侵袭、增殖活性降低。结论 ZEB23′UTR可能通过调控miR-200a/b/c的表达进而影响其对靶基因的转录后调控，增加细胞的侵袭、迁移能力，导致GES-1细胞的恶性转化倾向。
목적：탐토E합결합자지단백（ZEB）2기인3′비번역구（3′UTR）전염인위점막상피세포GES-1후대기증식、침습、천이적영향。방법장인공합성적ZEB23′UTR질립급miR-200b micmics채용지질체Lipofectamine 2000전염GES-1세포。분별설대조조、돌변조화ZEB23′UTR조，qRT-PCR검측전염후miR-200a/b/c급ZEB1/ZEB2 mRNA적표체。재설대조조、ZEB23′UTR조、ZEB23′UTR+무의서렬조화ZEB23′UTR+miR-200b micmics조。West-ern blot검측전염후ZEB1/ZEB2、기질금속단백매（MMP）-2/9、증식세포핵항원（PCNA）단백적표체；Transwell침습실험화화흔실험검측세포침습천이능력；MTT법검측세포증식활성。결과여대조조급돌변조상비，전염ZEB23′UTR조miR-200a/b/c적표체균하조，이miR-200b최위명현，ZEB1/ZEB2 mRNA급단백수평표체상조，기세포천이、침습、증식활성균증강（P<0.05）。ZEB23′UTR+miR-200b micmics조교ZEB23′UTR조천이、침습、증식활성강저。결론 ZEB23′UTR가능통과조공miR-200a/b/c적표체진이영향기대파기인적전록후조공，증가세포적침습、천이능력，도치GES-1세포적악성전화경향。
Objective To explore the effects of zinc finger E-box binding protein (ZEB)2 3′UTR gene transfection on proliferation, invasion and migration in human gastric epithelial cell line GES-1. Methods The synthetic ZEB2 3′UTR and miR-200b micmics were transfected into GES-1 cell line by lipofectamine 2000. We set up control grop, the mutation group and ZEB2 3′UTR group. Real-time quantitative PCR was performed to evaluate the expression levels of miR-200a/b/c and ZEB1/ZEB2 mRNAs after transfection.And then we set up control group, ZEB2 3′UTR group, ZEB2 3′UTR+negative control group and ZEB2 3′UTR+miR-200b micmics group. The protein expression levels of ZEB1, ZEB2, matrix metallopro-teinases (MMP) 2/9 and proliferating cell nuclear antigen (PCNA) were detected by Western blot assay. The invasion and mi-gration 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 de-creased, especially for miR-200b. And the expressions of ZEB1/ZEB2 were significantly increased at both mRNA and pro-tein levels after transfected with the ZEB2 3′UTR, enhancing the capability of migration,invasion,and proliferation (P <0.05). Compared with ZEB2 3′UTR group, the capabilities of proliferation,invasion and migration were significantly lower in combined group. Conclusion ZEB2 3′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-1 cells.