Transfection and Lentiviral Transduction

The transfection was performed when they were 60~80% confluent. The cells were transfected with the miR-320-5p/NC mimic or miR-320-5p/NC inhibitor (Ribobio, Guangzhou, China) using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The circHECTD1 or SLC2A1 was subcloned into the lentivirus vector ((polyA-MCS-UBI)RV-SV40-EGFP-IRES-puromycin) to construct LV- circHECTD1 vector, while LV-NC was used as the negative control, and (hU6-MCS-Ubiquitin-EGFP-IRES-puromycin) to construct sh-circHECTD1 or sh-SLC2A1 vector, while sh-NC was used as the negative control. All lentiviruses were constructed by Genechem (Shanghai, China). After the lentiviruses were cultured for 48 h, the lentiviruses were removed and replaced with fresh medium. The cells were then screened by culturing in the presence of 2 μg/mL puromycin after transfection.