Department of Gynecology and Obstetrics, West China Second University Hospital, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
Key Laboratory of Obstetrics and Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second Hospital, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
BMC Cancer. 2019 Jul 18;19(1):709. doi: 10.1186/s12885-019-5775-1.
The clinical behaviors and cytogenetics of solitary uterine leiomyomas (SUL) and multiple uterine leiomyomas (MUL) vary, which greatly affects the choice of treatments for reproductive-aged patients with leiomyomas. Our previous study demonstrated that a series of microRNAs, including miR-146b-5p, are dysregulated and play important roles in the development of SUL and MUL. Long non-coding RNAs (lncRNAs) can participate in the pathogenesis of several diseases by regulating the expression of microRNAs; however, their roles in regulating miR-146b-5b and in the pathology of leiomyomas are unclear.
Pair-matched uterine leiomyoma and adjacent normal myometrium tissue samples were collected from 37 patients with leiomyomas, including 15 with SUL and 22 with MUL. Six paired samples (three SUL and three MUL samples) were used for lncRNAs microarray analysis. Targeted lncRNAs were selected by bioinformatics analysis, and were verified by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and a dual-luciferase reporter assay. Growth curve analysis and qRT-PCR were used to evaluate the effect of silencing the lncRNA lnc-AL445665.1-4 on cell proliferation and miR-146b-5p expression, respectively.
There were 245 up-regulated and 243 down-regulated lncRNAs in SUL, and 119 up-regulated and 447 down-regulated lncRNAs in MUL. Fifty-five of the selected lncRNAs were predicted to target miR-146b-5p, which is up-regulated in SUL and down-regulated in MUL. Four lncRNAs were selected after Venn diagram analysis showing common dysregulation in the three groups. Lnc-AL445665.1-4 was selected for further exploration. qRT-PCR showed that lnc-AL445665.1-4 expression was significantly up-regulated in MUL compared with SUL in an additional 12 and 19 paired SUL-normal and MUL-normal samples, respectively. The dual-luciferase reporter assay demonstrated the presence of binding sites on lnc-AL445665.1 for miR-146b-5p. Silencing lnc-AL445665.1-4 not only inhibited cell proliferation but also negatively regulated the expression of miR-146b-5p.
Our results suggest that lnc-AL445665.1-4 may be involved in the development of MUL by interacting with miR-146b-5p. Further investigation of the roles of lncRNAs and miRNAs may help to optimize the clinical management of leiomyoma patients. Lnc-AL445665.1-4 could be a novel target for genetic therapy or serve as a biomarker for predicting the recurrence of MUL in patients that have undergone myomectomy.
单发子宫肌瘤（SUL）和多发子宫肌瘤（MUL）的临床行为和细胞遗传学不同，这极大地影响了生殖期子宫肌瘤患者治疗方法的选择。我们之前的研究表明，一系列 microRNAs，包括 miR-146b-5p，失调并在 SUL 和 MUL 的发生发展中发挥重要作用。长链非编码 RNA（lncRNA）可以通过调节 microRNAs 的表达参与多种疾病的发病机制；然而，它们在调节 miR-146b-5p 和子宫肌瘤发病机制中的作用尚不清楚。
收集 37 例子宫肌瘤患者配对的子宫肌瘤和邻近正常子宫肌层组织标本，包括 15 例 SUL 和 22 例 MUL。对 6 对配对样本（3 对 SUL 和 3 对 MUL 样本）进行 lncRNA 微阵列分析。通过生物信息学分析选择靶向 lncRNA，并通过定量逆转录-聚合酶链反应（qRT-PCR）和双荧光素酶报告基因检测进行验证。生长曲线分析和 qRT-PCR 分别用于评估沉默 lncRNA lnc-AL445665.1-4 对细胞增殖和 miR-146b-5p 表达的影响。
SUL 中有 245 个上调和 243 个下调的 lncRNA，MUL 中有 119 个上调和 447 个下调的 lncRNA。选择了 55 个 lncRNA 作为预测靶标 miR-146b-5p，miR-146b-5p 在 SUL 中上调，在 MUL 中下调。Venn 图分析显示三组共有 4 个 lncRNA 共同失调，选择 lnc-AL445665.1-4 进行进一步研究。qRT-PCR 显示，在另外 12 对和 19 对 SUL-正常和 MUL-正常配对样本中，MUL 中 lnc-AL445665.1-4 的表达明显高于 SUL。双荧光素酶报告基因检测表明 lnc-AL445665.1 上存在 miR-146b-5p 的结合位点。沉默 lnc-AL445665.1-4 不仅抑制细胞增殖，还负调控 miR-146b-5p 的表达。
我们的结果表明，lnc-AL445665.1-4 可能通过与 miR-146b-5p 相互作用参与 MUL 的发生发展。进一步研究 lncRNA 和 microRNA 的作用可能有助于优化子宫肌瘤患者的临床管理。lnc-AL445665.1-4 可能是一种新的遗传治疗靶点，也可以作为预测子宫肌瘤患者接受子宫肌瘤切除术复发的标志物。
