Department of Obstetrics and Gynecology, Affiliated Jiangyin Hospital of South-East University, Jiangyin 214400, China.
Department of Women Health Care, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210004, China.
Biomed Pharmacother. 2015 Oct;75:226-33. doi: 10.1016/j.biopha.2015.07.039. Epub 2015 Aug 24.
Our previous study showed that the expression of miR-197 in leiomyoma was down-regulated compared with myometrium. Further, miR-197 has been identified to affect uterine leiomyoma cell proliferation, apoptosis, and metastasis ability, though the responsible molecular mechanism has not been well elucidated. In this study, we sought to determine the expression patterns of miR-197 targeted genes and to explore their potential functions, participating Pathways and the networks that are involved in the biological behavior of human uterine leiomyoma.
After transfection of human uterine leiomyoma cells with miR-197, we confirmed the expression level of miR-197 using quantitative real-time PCR (qRT-PCR), and we detected the gene expression profiles after miR-197 over-expression through DNA microarray analysis. Further, we performed GO and Pathway analysis. The dominantly dys-regulated genes, which were up- or down-regulated by more than 10-fold, compared with parental cells, were confirmed using qRT-PCR technology.
Compared with the control group, miR-197 was up-regulated by 30-fold after miR-197 lentiviral transfection. The microarray data showed that 872 genes were dys-regulated by more than 2-fold in human uterine leiomyoma cells after miR-197 overexpression, including 537 up-regulated and 335 down-regulated genes. The GO analysis indicated that the dys-regulated genes were primarily involved in response to stimuli, multicellular organ processes, and the signaling of biological progression. Further, Pathway analysis data showed that these genes participated in regulating several signaling Pathways, including the JAK/STAT signaling Pathway, the Toll-like receptor signaling Pathway, and cytokine-cytokine receptor interaction. The qRT-PCR results confirmed that 17 of the 66 selected genes, which were up- or down-regulated more than 10-fold by miR-197, were consistent with the microarray results, including tumorigenesis-related genes, such as DRT7, SLC549, SFMBT2, FLJ37956, FBLN2, C10orf35, HOXD12, CACNG7, and LOC100134279.
Our study explored gene expression patterns after miR-197 overexpression and confirmed 17 dominantly dys-regulated genes, which could expand the insights into the function of miR-197 and the molecular mechanisms during the development and progression of uterine leiomyomas. This study might afford new clues for understanding the pathogenesis of uterine leiomyomas, and it could likely provide a unique method for diagnosing or predicting prognosis in the clinical treatment of leiomyoma.
我们之前的研究表明，与子宫肌层相比，子宫肌瘤中 miR-197 的表达下调。此外，已经确定 miR-197 影响子宫平滑肌瘤细胞的增殖、凋亡和转移能力，尽管其负责的分子机制尚未得到很好的阐明。在这项研究中，我们试图确定 miR-197 靶向基因的表达模式，并探讨它们的潜在功能、参与的途径以及涉及人类子宫肌瘤生物学行为的网络。
用 miR-197 转染人子宫肌瘤细胞后，我们使用定量实时 PCR(qRT-PCR)证实 miR-197 的表达水平，并通过 DNA 微阵列分析检测 miR-197 过表达后的基因表达谱。进一步进行了 GO 和途径分析。与亲本细胞相比，倍数变化超过 10 倍的明显上调或下调的主导失调基因，通过 qRT-PCR 技术进行了验证。
与对照组相比，miR-197 转染后 miR-197 的表达水平上调了 30 倍。微阵列数据显示，miR-197 过表达后，人子宫肌瘤细胞中 872 个基因失调超过 2 倍，其中 537 个上调，335 个下调。GO 分析表明，失调基因主要参与对刺激的反应、多细胞器官过程和生物进展的信号。此外，途径分析数据表明，这些基因参与调节几个信号通路，包括 JAK/STAT 信号通路、Toll 样受体信号通路和细胞因子-细胞因子受体相互作用。qRT-PCR 结果证实，在 miR-197 上调或下调超过 10 倍的 66 个选定基因中，有 17 个与微阵列结果一致，包括与肿瘤发生相关的基因，如 DRT7、SLC549、SFMBT2、FLJ37956、FBLN2、C10orf35、HOXD12、CACNG7 和 LOC100134279。
我们的研究探讨了 miR-197 过表达后的基因表达模式，并证实了 17 个明显失调的基因，这可能扩展了对 miR-197 功能和子宫平滑肌瘤发生和进展过程中分子机制的认识。这项研究可能为理解子宫肌瘤的发病机制提供新的线索，并可能为临床治疗平滑肌瘤的诊断或预测预后提供独特的方法。
