Novel insights into the circ_0003489/let-7b-5p/GLUT1 axis and its possible role in multiple myeloma

Background: Circular RNAs (circRNAs) act as vital players in multiple myeloma (MM). Herein, we focused on the function of hsa_circ_0003489 (circ_0003489) in MM development and bortezomib (BTZ) resistance.

Methods: Relative RNA levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Relative protein levels were evaluated by Western blotting or immunohistochemistry (IHC). The 5'-ethynyl-2'-deoxyuridine (EdU) and cell colony formation (CF) assays were conducted for cell proliferation. Cell counting kit-8 assay was used to evaluate the BTZ resistance. Flow cytometry analysis was performed for cell apoptosis analysis. Glycolysis was determined by detecting the levels of ECAR, glucose consumption, and lactate production. Dual-luciferase reporter and RNA pull-down assays were carried out to analyze the relationships of circ_0003489 with let-7b-5p microRNA and glucose transporter 1 (GLUT1) glucose transporter protein. Xenograft models were conducted to assess the function of circ_0003489 in vivo.

Results: Indeed, as shown by qRT-PCR, bone marrow samples of MM patients showed an upregulation of circ_0003489 RNA in comparison to normal controls (P < 0.0001). In in vitro experiments in MM cells, silencing of circ_0003489 repressed cell proliferation, BTZ resistance, and glycolysis. Furthermore, blocking circ_0003489 facilitated in vitro the apoptosis of MM cells. In vivo experiments showed that silencing circ_0003489 decreased tumor formation. Signaling experiments demonstrated that circ_0003489 sponged let-7b-5p microRNA and negatively regulated let-7b-5p microRNA expression. Loss of let-7b-5p microRNA ameliorated circ_0003489 silencing-mediated effects on MM cell malignant behaviors and BTZ resistance. Moreover, we showed that GLUT1 glucose transporter was targeted by let-7b-5p mircoRNA. GLUT1 enhancement reversed the repressive impacts of let-7b-5p upregulation on MM cell malignant behaviors and BTZ resistance.

Conclusion: We suggest that circ_0003489 RNA knockdown inhibited MM progression and reversed BTZ-induced resistance of MM growth by let-7b-5p microRNA regulated function of GLUT1 glucose transporter.