Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
Nucleic Acid Ther. 2013 Feb;23(1):29-34. doi: 10.1089/nat.2012.0385. Epub 2012 Dec 3.
MicroRNAs (miRNAs) play important roles in tumorigenesis and metastasis. In this study, we investigated miR-200b expression in endometrial adenocarcinomas and normal adjacent tissues and found that miR-200b is more highly expressed in cancer tissues than in normal adjacent tissues. A novel target of miR-200b, tissue inhibitor of metalloproteinase 2 (TIMP2), was predicted using a bioinformatics approach and was confirmed in human endometrial cancer cell line HEC-1A cells by luciferase assay, quantitative real-time polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay. We found that miR-200b repressed TIMP2 expression at both the messenger RNA and protein levels, although a family member, miR-200a, had no such effect. Using reverse gelatin zymography, we showed that miR-200b enhances matrix metallopeptidase 2 (MMP2) activity by downregulating TIMP2 expression in HEC-1A cells. These data suggest that miR-200b may play an important role in the metastasis of endometrial adenocarcinomas.
微小 RNA(miRNAs)在肿瘤发生和转移中发挥重要作用。在这项研究中,我们调查了 miR-200b 在子宫内膜腺癌和正常相邻组织中的表达,发现 miR-200b 在癌症组织中的表达高于正常相邻组织。通过生物信息学方法预测了 miR-200b 的一个新靶标,组织金属蛋白酶抑制剂 2(TIMP2),并通过荧光素酶测定、定量实时聚合酶链反应、western blot 和酶联免疫吸附试验在人子宫内膜癌细胞系 HEC-1A 细胞中得到证实。我们发现 miR-200b 在信使 RNA 和蛋白质水平上均抑制 TIMP2 的表达,尽管家族成员 miR-200a 没有这种作用。通过反转胶凝酶谱分析,我们表明 miR-200b 通过下调 HEC-1A 细胞中的 TIMP2 表达来增强基质金属蛋白酶 2(MMP2)的活性。这些数据表明 miR-200b 可能在子宫内膜腺癌的转移中发挥重要作用。