Department of Obstetrics and Gynecology, Key Laboratory of Maternal-Fetal Medicine of Liaoning Province, Key Laboratory of Obstetrics and Gynecology of Higher Education of Liaoning Province, Shengjing Hospital of China Medical University, 39 Huaxiang Road, Shenyang, 110021, People's Republic of China.
Cell Death Dis. 2018 Feb 21;9(3):303. doi: 10.1038/s41419-018-0291-9.
Long noncoding RNAs (lncRNAs) have been implicated in tumorigenesis and cancer progression and are tightly associated with the phenotypes of numerous cancers. However, the functional roles underlying these effects are unknown. The expression levels of LINC01016, miR-302a-3p, miR-3130-3p, NFYA, and SATB1 were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) in 33 endometrial cancer tissues and 20 normal tissues. Bioinformatics analyses, luciferase reporter analyses, chromatin immunoprecipitation (ChIP) assays, and qRT-PCR assays were performed to verify potential binding sites. The qRT-PCR and western blot were used to identify the regulatory mechanisms of LINC01016 in cell biological behavior, which were also examined by cell counting kit -8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) assays, flow cytometry, wound healing assays, and transwell assays. LINC01016 was substantially upregulated in endometrial cancer tissues, and LINC01016 silencing abolished the malignant behavior of endometrial cancer cells. LINC01016 positively rescued the downstream gene nuclear factor YA (NFYA) by competitively "sponging" miR-302a-3p and miR-3130-3p. In turn, these two miRNAs could inhibit LINC01016 transcription, thus forming two reciprocal repression cycles, which influenced the biological behavior of endometrial cancer cells. MiR-302a-3p and miR-3130-3p could specifically bind with the 3'-UTR regions of NFYA, and NFYA could upregulate the expression of special AT-rich sequence-binding protein 1 (SATB1) as a transcriptional factor. This study was the first to show that the LINC01016-miR-302a-3p/miR-3130-3p/NFYA/SATB1 axis played a crucial role in the occurrence of endometrial cancer. These findings may provide relevant insights into the diagnosis and therapy of endometrial cancer.
长链非编码 RNA（lncRNAs）已被牵涉到肿瘤发生和癌症进展中，并与许多癌症的表型密切相关。然而，这些影响背后的功能作用尚不清楚。通过实时定量聚合酶链反应（qRT-PCR）在 33 份子宫内膜癌组织和 20 份正常组织中评估 LINC01016、miR-302a-3p、miR-3130-3p、NFYA 和 SATB1 的表达水平。通过生物信息学分析、荧光素酶报告分析、染色质免疫沉淀（ChIP）分析和 qRT-PCR 分析验证潜在的结合位点。通过 qRT-PCR 和 Western blot 鉴定 LINC01016 对细胞生物学行为的调节机制，并通过细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）测定、流式细胞术、划痕愈合测定和 Transwell 测定进行检测。LINC01016 在子宫内膜癌组织中明显上调，沉默 LINC01016 可消除子宫内膜癌细胞的恶性行为。LINC01016 通过竞争性“海绵”miR-302a-3p 和 miR-3130-3p 正向挽救下游基因核因子 YA（NFYA）。反过来，这两个 miRNA 可以抑制 LINC01016 的转录，从而形成两个相互抑制的循环，影响子宫内膜癌细胞的生物学行为。miR-302a-3p 和 miR-3130-3p 可以特异性结合 NFYA 的 3'-UTR 区域，NFYA 作为转录因子可以上调特殊富含 AT 序列结合蛋白 1（SATB1）的表达。这项研究首次表明，LINC01016-miR-302a-3p/miR-3130-3p/NFYA/SATB1 轴在子宫内膜癌的发生中起着关键作用。这些发现可能为子宫内膜癌的诊断和治疗提供相关见解。
