Zhang Xiaona, Zhu Yanmei, Wu Jun-Dong, Zhou Yanchun, Chen Weibing, Gu Wei
Department of Pathophysiology, The Key Immunopathology Laboratory of Guangdong Province, Shantou University Medical College, Shantou, Guangdong Province, 515041, China.
Tumor Hospital, Shantou University Medical College, Shantou, Guangdong Province, 515041, China.
Noncoding RNA Res. 2022 Jul 4;7(3):164-170. doi: 10.1016/j.ncrna.2022.06.003. eCollection 2022 Sep.
Increasing studies have shown that lncRNAs often play roles through interaction with miRNAs to control gene expression by inhibiting translation or facilitating degradation of target mRNAs. Here, we report that two lncRNAs, MACC1-AS1 and UCA1 are coordinately expressed in breast cancer cells and share the ability to interact with multiple miRNAs to mediate the expression of different genes.
Targetscan, starBase and miRDB databases were used to predict the relationships of MACC1-AS1/UCA1-miRNA-mRNA network. qRT-PCR, and RNA sequencing were used to study the differential expression of lncRNAs and miRNA-targeted genes in breast cancer cells. RIP, RNA pull-down and luciferase assays were performed to confirm the molecular interactions of MACC1-AS1 or UCA1 with predicted miRNAs. The role of lncRNA-mediated miRNA-mRNA interactions in cell proliferation was examined by MTT assays following loss-of-function and gain-of-function effects.
We identified a lncRNA-miRNA-mRNA regulatory network in breast cancer cells, in which a number of mRNAs can be co-regulated by MACC1-AS1 and UCA1 lncRNAs. Each lncRNA possesses the capacity as a ceRNA to compete with various mRNA-targeting miRNAs. Interaction of MACC1-AS1 or UCA1 with individual miRNAs is able to increase the expression of the same target mRNAs, such as TBL1X and MEF2D, thus affecting cancer-cell growth phenotype.
Our study suggests that in each cell type, there is a balance of interactions between certain lncRNAs and miRNAs. Disrupting the balance would eventually affect the expression of miRNA-targeted genes and cell proliferation.
越来越多的研究表明，长链非编码RNA（lncRNAs）常通过与微小RNA（miRNAs）相互作用来发挥作用，通过抑制翻译或促进靶标信使核糖核酸（mRNAs）的降解来控制基因表达。在此，我们报告两种lncRNAs，即MACC1-AS1和UCA1，在乳腺癌细胞中协同表达，并具有与多种miRNAs相互作用以介导不同基因表达的能力。
使用Targetscan、starBase和miRDB数据库预测MACC1-AS1/UCA1-miRNA-mRNA网络的关系。采用定量逆转录聚合酶链反应（qRT-PCR）和RNA测序研究lncRNAs和miRNA靶向基因在乳腺癌细胞中的差异表达。进行RNA免疫沉淀（RIP）、RNA下拉和荧光素酶测定以证实MACC1-AS1或UCA1与预测的miRNAs之间的分子相互作用。在功能丧失和功能获得效应后，通过MTT测定检查lncRNA介导的miRNA-mRNA相互作用在细胞增殖中的作用。
我们在乳腺癌细胞中鉴定出一个lncRNA-miRNA-mRNA调控网络，其中许多mRNAs可由MACC1-AS1和UCA1 lncRNAs共同调控。每个lncRNA都具有作为竞争性内源RNA（ceRNA）与各种靶向mRNA的miRNAs竞争的能力。MACC1-AS1或UCA1与单个miRNAs的相互作用能够增加相同靶标mRNAs（如TBL1X和MEF2D）的表达，从而影响癌细胞生长表型。
我们的研究表明，在每种细胞类型中，某些lncRNAs和miRNAs之间的相互作用存在平衡。破坏这种平衡最终会影响miRNA靶向基因的表达和细胞增殖。
