Long Dianbo, Xu Yiyang, Mao Guping, Xin Ruobing, Deng Zengfa, Liao Hongyi, Li Zhiwen, Yang Zhi, Yu Baoxi, Yang Zhijian, He Aishan, Zhang Ziji, Kang Yan
Department of Joint Surgery, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, 510080, China.
Guangdong Provincial Key Laboratory of Orthopedics and Traumatology, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, 510080, China.
Cell Death Discov. 2022 Jan 10;8(1):19. doi: 10.1038/s41420-021-00806-4.
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.
转运RNA衍生片段（tRFs）是新型非编码RNA，近期研究表明，tRNA和tRFs通过基因表达的转录后调控在细胞代谢中发挥重要作用。然而，tRFs是否调节前交叉韧带（ACL）的细胞代谢仍不清楚。本研究旨在探讨tRFs在ACL细胞代谢中的作用及作用机制。使用tRF阵列确定不同人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确定TRF365的表达。用TRF365模拟物或TRF365抑制剂转染ACL细胞，以确定TRF365是否调节IKBKB的表达。进行了拯救实验和双荧光素酶报告基因检测，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细胞中表达较弱。IKBKB在OA ACL和白细胞介素-1β处理的ACL细胞中高表达；用TRF365模拟物转染可抑制IKBKB的表达，而用TRF365抑制剂转染则产生相反的效果。双荧光素酶报告基因检测表明，TRF365通过与其3'-UTR结合使IKBKB的表达沉默。因此，TRF365通过靶向IKBKB调节ACL细胞的代谢。总之，TRF365可能为ACL退变及OA病理生理过程的研究提供新的方向。
