tRNA-derived fragment TRF365 regulates the metabolism of anterior cruciate ligament cells by targeting IKBKB.
tRNA-derived fragment TRF365 regulates the metabolism of anterior cruciate ligament cells by targeting IKBKB.
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tRNA衍生片段TRF365通过靶向IKBKB调节前十字韧带细胞的代谢
DOI：
10.1038/s41420-021-00806-4
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发表时间：
2022-01-10
影响因子：
7
通讯作者：
Kang Y
中科院分区：
文献类型：
--
作者：
Long D;Xu Y;Mao G;Xin R;Deng Z;Liao H;Li Z;Yang Z;Yu B;Yang Z;He A;Zhang Z;Kang Y
tRNA衍生片段（tRF）是一种新的非编码RNA，最近的研究表明，tRNA和tRF通过基因表达的转录后调控在细胞代谢中具有重要功能。然而，tRF 是否调节前十字韧带 (ACL) 的细胞代谢仍不清楚。本研究的目的是探讨tRFs在ACL细胞代谢中的作用和作用机制。使用tRF阵列测定不同人ACL细胞中的tRF表达谱，并使用定量实时聚合酶链反应和荧光原位杂交测定TRF365表达。用 TRF365 模拟物或 TRF365 抑制剂转染 ACL 细胞，以确定 TRF365 是否调节 IKBKB 表达。进行救援实验和双荧光素酶报告基因测定以确定 IKBKB 的 3'-非翻译区 (UTR) 是否具有 TRF365 结合位点。 TRF365 在骨关节炎 (OA) ACL 和白细胞介素 1β 处理的 ACL 细胞中弱表达。 IKBKB在OA ACL和IL-1β处理的ACL细胞中高表达；用TRF365模拟物转染抑制IKBKB表达，而用TRF365抑制剂转染则具有相反的效果。双荧光素酶报告基因检测显示，TRF365 通过与其 3'-UTR 结合来沉默 IKBKB 的表达。因此，TRF365通过靶向IKBKB来调节ACL细胞的代谢。综上所述，TRF365可能为ACL变性和OA病理生理过程的研究提供新的方向。
tRNA-derived fragments (tRFs) are new noncoding RNAs, and recent studies have shown that tRNAs and tRFs have important functions in cell metabolism via posttranscriptional regulation of gene expression. However, whether tRFs regulate cellular metabolism of the anterior cruciate ligament (ACL) remains elusive. The aim of this study was to investigate the role and action mechanism of tRFs in ACL cell metabolism. A tRF array was used to determine tRF expression profiles in different human ACL cells, and quantitative real-time polymerase chain reaction and fluorescence in situ hybridisation were used to determine TRF365 expression. ACL cells were transfected with a TRF365 mimic or a TRF365 inhibitor to determine whether TRF365 regulates IKBKB expression. A rescue experiment and dual-luciferase reporter assay were conducted to determine whether the 3′-untranslated region (UTR) of IKBKB has a TRF365-binding site. TRF365 was weakly expressed in osteoarthritis (OA) ACL and interleukin-1β-treated ACL cells. IKBKB was highly expressed in OA ACL and interleukin-1β-treated ACL cells; transfection with the TRF365 mimic suppressed IKBKB expression, whereas transfection with the TRF365 inhibitor had the opposite effect. A dual-luciferase reporter assay showed that TRF365 silenced the expression of IKBKB by binding to its 3′-UTR. Thus, TRF365 regulates the metabolism of ACL cells by targeting IKBKB. In summary, TRF365 may provide a new direction for the study of ACL degeneration and on the pathophysiological process of OA.
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