高通量测序分析分化型甲状腺癌中 SREBP1 相关差异表达 microRNA。

Department of Clinical Pharmacology, Xiangya Hospital, Central South University, Changsha, P.R. China.

Institute of Clinical Pharmacology, Central South University and Hunan Key Laboratory of Pharmacogenetics, Changsha, P.R. China.

Anticancer Res. 2023 Oct;43(10):4435-4446. doi: 10.21873/anticanres.16639.

BACKGROUND/AIM: MicroRNAs (miRNAs) interact with mRNAs and play important roles in progression and prognosis in multiple cancers. Sterol regulatory element-binding protein 1 (SREBP1) is an important lipid metabolism regulatory gene. The aim of the present study was to analyze the profiles of miRNAs that are associated with SREBP1 expression in differentiated thyroid carcinoma (DTC).

MATERIALS AND METHODS

In the present study, a high-throughput small RNA sequencing (miRNA-Seq) method was used to investigate differences in miRNA profiling with versus without interference with SREBP1 expression via small interfering RNA. Real-time qPCR (qRT-PCR) was performed to confirm the results.

RESULTS

A total of 1,393 conserved and 84 novel miRNAs were successfully discovered. In two separate batches, a total of 27 differentially expressed miRNAs (11 up-regulated and 16 down-regulated) were observed in BCPAP cells after SREBF1 interference with two distinct siRNA fragments, as compared to the control siRNA treatment. Hsa-miR-941, hsa-miR-27a-5p, hsa-miR-29a-3p, hsa-miR-100-5p, and hsa-miR-21-3p were selected for validation using qRT-PCR. The qRT-PCR results were consistent with the sequencing data. Gene Ontology enrichment showed that the predicted targets of these miRNAs were mainly involved in the regulation of system development, metabolism and protein binding cellular processes, and metabolic processes. Kyoto Encyclopedia of Genes and Genomes pathways analysis showed that the predicted target genes were involved in several signaling pathways, including the Ras, MAPK, insulin, thyroid hormone, and metabolic pathway signaling pathways.

CONCLUSION

Differentially expressed miRNAs and their target genes may play an important role in the progression and prognosis of DTC that is associated with SREBP1 expression.

背景/目的：微小 RNA（miRNA）与 mRNAs 相互作用，在多种癌症的进展和预后中发挥重要作用。固醇调节元件结合蛋白 1（SREBP1）是一种重要的脂质代谢调节基因。本研究旨在分析与分化型甲状腺癌（DTC）中 SREBP1 表达相关的 miRNA 谱。

材料和方法

本研究采用高通量小 RNA 测序（miRNA-Seq）方法，通过小干扰 RNA 干扰 SREBP1 表达，研究 miRNA 谱的差异。采用实时 qPCR（qRT-PCR）进行验证。

结果

成功发现了 1393 个保守 miRNA 和 84 个新 miRNA。在两个独立批次中，与对照 siRNA 处理相比，用两种不同的 siRNA 片段干扰 SREBF1 后，BCPAP 细胞中共有 27 个差异表达 miRNA（11 个上调和 16 个下调）。使用 qRT-PCR 验证了 hsa-miR-941、hsa-miR-27a-5p、hsa-miR-29a-3p、hsa-miR-100-5p 和 hsa-miR-21-3p。qRT-PCR 结果与测序数据一致。基因本体富集分析显示，这些 miRNA 的预测靶基因主要参与系统发育、代谢和蛋白质结合的细胞过程以及代谢过程的调节。京都基因与基因组百科全书通路分析显示，预测靶基因参与了几个信号通路，包括 Ras、MAPK、胰岛素、甲状腺激素和代谢途径信号通路。