The interaction between miR-148a and DNMT1 suppresses cell migration and invasion by reactivating tumor suppressor genes in pancreatic cancer
The interaction between miR-148a and DNMT1 suppresses cell migration and invasion by reactivating tumor suppressor genes in pancreatic cancer
复制标题
miR-148a和DNMT1之间的相互作用通过重新激活胰腺癌中的抑癌基因来抑制细胞迁移和侵袭
DOI：
10.3892/or.2018.6700
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发表时间：
2018-11-01
期刊：
影响因子：
4.2
通讯作者：
Xiao, Weidong
中科院分区：
文献类型：
--
作者：
Hong, Le;Sun, Gen;Xiao, Weidong
DNA甲基化是细胞用来控制基因表达的表观遗传机制，在肿瘤发生中起重要作用。DNA甲基转移酶1（DNMT 1）负责维持DNA甲基化模式。在包括胰腺癌在内的许多恶性肿瘤中观察到DNMT 1的过表达，并导致几个关键肿瘤抑制基因（TSGs）的沉默。最近的研究表明，microRNAs（miRNAs/miRs）参与了DNMT 1的表达调控，而DNMT 1过表达引起的启动子甲基化与某些miRNAs功能障碍有关。本研究旨在揭示miR-148 a与DNMT 1的相互作用及其对胰腺癌细胞增殖、迁移和侵袭的影响。首先，通过逆转录-定量聚合酶链反应（PCR）检测胰腺癌组织和AsPC-1细胞中DNMT 1和miR-148 a的表达水平。第二，采用甲基化特异性PCR方法检测DNMT 1对miR-148 a的调控作用。进一步利用生物信息学分析和双荧光素酶报告基因分析验证miR-148 a与DNMT 1的靶位关系。最后，进行体外挽救实验以评估miR-148 a对AsPC-1细胞中TSGs表达和恶性表型的影响。结果表明，DNMT 1在胰腺癌中异常上调，并负责miR-148 a启动子的高甲基化。此外，双荧光素酶报告基因检测显示DNMT 1是miR-148 a的直接靶点，miR-148 a的恢复可以通过靶向DNMT 1激活AsPC-1胰腺癌细胞系中的TSGs，如p16、前脑啡肽原和Ras相关结构域家族成员1。这些结果表明，在胰腺癌中miR-148 a和DNMT 1之间存在相互作用。值得注意的是，miR-148 a过表达显著抑制AsPC-1细胞的细胞增殖、迁移和侵袭。因此，miR-148 a可能成为胰腺癌治疗的新靶点。
DNA methylation is an epigenetic mechanism that cells use to control gene expression, which serves an important role in tumorigenesis. DNA methyltransferase 1 (DNMT1) is responsible for the maintenance of the pattern of DNA methylation. Overexpression of DNMT1 is observed in numerous malignant tumors, including pancreatic cancer, and results in silencing of several key tumor suppressor genes (TSGs). Recent studies have suggested that microRNAs (miRNAs/miRs) contribute to the regulation of DNMT1 expression, and promoter hypermethylation caused by DNMT1 overexpression is associated with the dysfunction of some miRNAs. The present study aimed to reveal the interaction between miR-148a and DNMT1, and its effects on cell proliferation, migration and invasion of pancreatic cancer cells. Initially, the expression levels of DNMT1 and miR-148a were detected in pancreatic cancer tissues and AsPC-1 cells by reverse transcription-quantitative polymerase chain reaction (PCR). Secondly, the regulatory effects of DNMT1 on miR-148a were evaluated using methylation-specific PCR. Furthermore, bioinformatics analysis and dual luciferase reporter assay were used to verify the target relationship between miR-148a and DNMT1. Finally, in vitro rescue experiments were conducted to evaluate the effects of miR-148a on the expression of TSGs and the malignant phenotype in AsPC-1 cells. The results demonstrated that DNMT1 was aberrantly upregulated in pancreatic cancer, and was responsible for hypermethylation of the miR-148a promoter. Furthermore, DNMT1 was revealed as a direct target of miR-148a by dual luciferase reporter assay, and restoration of miR-148a could reactivate TSGs, such as p16, preproenkephalin and Ras association domain family member 1 by targeting DNMT1 in the AsPC-1 pancreatic cancer cell line. These results indicated that an interaction exists between miR-148a and DNMT1 in pancreatic cancer. Notably, miR-148a overexpression significantly inhibited cell proliferation, migration and invasion in AsPC-1 cells. Therefore, miR-148a may serve as a novel therapeutic target for the treatment of pancreatic cancer.