5'-isomiR 是 miR-1246（一种肺癌的候选生物标志物）在血清中最丰富的序列。,Molecular Medicine Reports

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5'-isomiR 是 miR-1246（一种肺癌的候选生物标志物）在血清中最丰富的序列。
Molecular Medicine Reports ( IF 3.4 ) Pub Date : 2023-03-24 , DOI: 10.3892/mmr.2023.12979
Toshiko Aiso ₁ , Makiko Ueda ₁

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MicroRNA (miRNA/miR) 5'-isoforms (5'-isomiRs) 与 microRNA 数据库中注册的规范序列的 5' 末端长度不同。与目标 mRNA 结合的 miRNA 的“种子序列”是从 5' 端开始的 2-8 个核苷酸；因此，5' 端的移位会导致“种子移位”。从 miRNA 深度测序中积累的数据表明，在大量 miRNA 中，对应于特定异构体的序列，而不是规范形式，是最丰富的。迄今为止，研究主要集中在循环 miRNA 作为标记物或细胞间通讯因子。miR-1246 在血清中含量丰富，是食管鳞状细胞癌、胰腺癌、肝细胞癌的候选诊断和预后标志物，结直肠腺癌和非小细胞肺癌 (NSCLC)。本研究通过片段分析对血清 miR-1246 的 5'-末端进行分析，发现比标准序列短两个碱基的 5'-isomiR 是 NSCLC 患者和健康供体中最丰富的序列. 为了量化 5'-isomiR，使用了基于等位基因特异性 PCR 引物的 5'-isomiR 特异性引物，主要是因为用于 miRNA 逆转录定量 PCR 的商业可用方法无法区分序列，尤其是位于 5' 的序列miRNA 的末端。NSCLC患者的总miR-1246水平显着升高；相比之下，规范序列的水平显着降低。在总 miR-1246 水平和 5'-isomiR 水平之间观察到显着的正相关，但与规范序列的水平不相关。这些结果意味着 NSCLC 患者血清 miR-1246 水平的增加取决于 5'-isomiR 的增加。

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5'‑isomiR is the most abundant sequence of miR‑1246, a candidate biomarker of lung cancer, in serum.

MicroRNA (miRNA/miR) 5'‑isoforms (5'‑isomiRs) differ from canonical sequences registered in the microRNA database in the length of their 5' ends. The 'seed sequence' of miRNAs that bind to target mRNAs is 2‑8 nucleotides from the 5' end; thus, shifts at the 5' end can cause a 'seed shift'. Accumulating data from miRNA deep sequencing have revealed that, in a substantial number of miRNAs, sequences corresponding to specific isomiRs, not the canonical form, are the most abundant. Studies have so far focused on circulating miRNAs as either markers or intercellular communication factors. miR‑1246 is abundant in the serum and is a candidate diagnostic and prognostic marker for esophageal squamous cell carcinoma, pancreatic cancer, hepatocellular carcinoma, colorectal adenocarcinoma and non‑small cell lung cancer (NSCLC). The present study analyzed the 5'‑end of serum miR‑1246 by fragment analysis and found that a 5'‑isomiR, which is two bases shorter than the canonical sequence, was the most abundant sequence in patients with NSCLC as well as healthy donors. To quantify the 5'‑isomiR, 5'‑isomiR‑specific primers based on primers for allele specific‑PCR were used, primarily because commercially available methods for miRNA Reverse transcription‑quantitative PCR cannot discriminate among sequences, especially those located at the 5' end of miRNA. The total miR‑1246 levels were significantly increased in patients with NSCLC; by contrast, the level of the canonical sequence was significantly decreased. Significant positive correlations were observed between the total miR‑1246 levels and the 5'‑isomiR levels, but not that of the canonical sequence. These results imply that the increase in levels of serum miR‑1246 in patients with NSCLC depends on increase of the 5'‑isomiR.

更新日期：2023-03-24

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