H22954, a long non-coding RNA, inhibits glucose uptake in leukemia cells in a GLUT10-dependent manner

To investigate the performance of H22954, a novel long non-coding RNA (lncRNA), in inhibiting glucose uptake in leukemia cells. ¹⁸F-FDG uptake, RNA half-life quantitative real-time polymerase chain reaction (qRT-PCR) and luciferase assays were performed to detect the glucose uptake in the condition of leukemia. Microarrays and qRT-PCR analyses were used to identify the related genes or proteins and elucidate the underlying these processes. H22954, a novel lncRNA, inhibited glucose uptake in leukemia cells. Using bioinformatics and microarray analyses, GLUT10 was identified as a possible target molecule of H22954. H22954 targeted the 3′untranslated region of GLUT10. In the luciferase assay, the luciferase activity of pGL3-GLUT10 was inhibited by H22954. Consistently, H22954 expression levels were inversely correlated with GLUT10 expression in cell lines and acute myeloid leukemia (AML) samples. Conversely, the degradation rate of <i>GLUT10</i> mRNA was increased after H22954 overexpression. Moreover, glucose uptake was recovered when the GLUT10-interaction sites in H22954 were mutated. The lncRNA H22954 regulated GLUT10 expression to inhibit glucose uptake in leukemia cells. Our findings provide potentially valuable data for designing new targeted strategies based on H22954.

本研究旨在探究新型长链非编码RNA（long non-coding RNA，lncRNA）H22954对白血病细胞葡萄糖摄取的抑制作用。实验通过检测¹⁸F-氟代脱氧葡萄糖（18F-FDG）摄取、RNA半衰期、实时定量聚合酶链反应（quantitative real-time polymerase chain reaction，qRT-PCR）及荧光素酶实验，分析白血病环境下的细胞葡萄糖摄取水平；并采用基因芯片及qRT-PCR分析，鉴定相关基因与蛋白，阐明上述调控过程的潜在分子机制。结果显示，新型lncRNA H22954可抑制白血病细胞的葡萄糖摄取。通过生物信息学及基因芯片分析，本研究鉴定出葡萄糖转运蛋白10（glucose transporter 10，GLUT10）为H22954的潜在靶分子；H22954可靶向结合GLUT10的3′非翻译区（3′untranslated region，UTR）。荧光素酶实验结果表明，H22954可抑制pGL3-GLUT10的荧光素酶活性。与此一致，在细胞系及急性髓系白血病（acute myeloid leukemia，AML）样本中，H22954的表达水平与GLUT10的表达呈负相关。反之，在H22954过表达后，GLUT10 mRNA的降解速率显著升高。此外，当H22954上与GLUT10结合的位点发生突变时，细胞的葡萄糖摄取水平得以恢复。综上，lncRNA H22954通过调控GLUT10的表达，抑制白血病细胞的葡萄糖摄取。本研究结果为基于H22954的新型靶向治疗策略的开发提供了具有潜在应用价值的实验依据。