Ruan Hongjie, Liang Xin, Zhao Wei, Ma Li, Zhao Yibing
Department of Gynecology, Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University, Nanjing 210000, China.
Department of Clinical Laboratory, Jiangsu Provincial Hospital of Traditional Chinese Medicine, Nanjing 210000, China.
Biomed Pharmacother. 2017 May;89:812-818. doi: 10.1016/j.biopha.2017.02.091. Epub 2017 Mar 6.
MiRNAs are known to play important roles in cancer cell development. However, the pattern and biological role of miR-183 in endometrial cancer (EC) have not been completely unexplored. Here, we found that miR-183 was upregulated in endometrial cancer cells. The purpose of the study was to evaluate the function of miR-183 in the endometrial cancer cell line and the mechanisms regulating its direct target protein in these processes. The mRNA and protein expressions were analyzed by quantitative RT-PCR and western blotting, respectively. The experiments about MTT assay, colony formation assay and transwell assay showed that miR-183 can positively regulate cell proliferation, migration and invasion in vitro. Furthermore, the in vivo experiments indicated that knockdown of miR-183 significantly attenuated EC cells growth. Mechanistically, luciferase reporter assay and western blotting assay was conducted to confirm target associations. The data analysis revealed that MMP-9 as a direct target of miR-183 in EC and there was a negatively relationship between miR-183 and MMP-9 expression in EC cells. Taken together, our results demonstrated that miR-183 plays a critical role in EC tumorigenesis and metastasis by suppressing MMP-9 expression, which may be an attractive therapeutic target for the treatment of endometrial cancer.
已知微小RNA(miRNAs)在癌细胞发育中发挥重要作用。然而,miR-183在子宫内膜癌(EC)中的模式和生物学作用尚未完全明晰。在此,我们发现miR-183在子宫内膜癌细胞中上调。本研究的目的是评估miR-183在子宫内膜癌细胞系中的功能以及在这些过程中调节其直接靶蛋白的机制。分别通过定量逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法分析mRNA和蛋白质表达。关于MTT法、集落形成试验和Transwell试验的实验表明,miR-183在体外可正向调节细胞增殖、迁移和侵袭。此外,体内实验表明,敲低miR-183可显著减弱EC细胞生长。从机制上讲,进行了荧光素酶报告基因试验和蛋白质免疫印迹试验以确认靶标关联。数据分析显示,基质金属蛋白酶-9(MMP-9)是EC中miR-183的直接靶标,并且在EC细胞中miR-183与MMP-9表达之间存在负相关关系。综上所述,我们的结果表明,miR-183通过抑制MMP-9表达在EC肿瘤发生和转移中起关键作用,这可能是治疗子宫内膜癌的一个有吸引力的治疗靶点。
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