Sh con

The pcDNA3.1 vector carrying SBF2-AS1 (SBF2-AS1), pcDNA3.1 vector carrying ADAM17 (ADAM17) or empty vector (vector) were obtained from RiboBio (Guangzhou, Guangdong, China). MiR-338-3p mimics, miRNA negative control (miR-NC), miR-338-3p inhibitor, miR-NC inhibitor, small interfering RNA (siRNA) targeting SBF2-AS1 or ADAM17 (si-SBF2-AS1, si-ADAM17) and scramble control siRNA (si-Con) were purchased from RiboBio. 2×10⁴ A549 or H1975 cells were transfected with 100 μmol/L miRNAs, or 5 μg pcDNA3.1, or 50 nM siRNA using 7.5 μl of Lipofectamine 3000 (Thermo Fisher Scientific) in 125 μl of Opti-MEMTM medium combination with 5 μl of p3000 for twenty-four hours. For functional assay in vivo, sh-SBF2-AS1 and sh-Con were purchased from RiboBio and constructed into A549 cell. The cDNA sequences of SBF2-AS1 were constructed by Shanghai Jima Pharmaceutical Technology Co., Ltd., (Shanghai, China) and were subcloned into pLKO.1 lentivirus vector (Addgene). pLKO.1 empty vector was used as negative control plasmid. Then, lentivirus plasmid was transfected into HEK-293T cell along with lentivirus packaging plasmids (psPAX2 and pMD2.G, Addgene). 72 hours after transfection, cell supernatants were collected and A549 cell was infected with lentivirus, followed by the screening of 1.5 μg/ml puromycin (Sigma). After 7 days, stable lentivirus transfected A549 cell lines were obtained.

The small hairpin RNAs against circSEPT9 (sh-circSEPT9, 5ʹ-GCCAGGAGGCCTTGAAAAGAT-3ʹ) and SLC1A5 (sh-SLC1A5, 5ʹ-TCAGCAGCCTTTCGCTCATACTCTA-3ʹ), miR-149-5p mimic (5ʹ-UCUGGCUCCGUGUCUUCACUCCC-3ʹ), miR-149-5p inhibitor (5ʹ-GGGAGUGAAGACACGGAGCCAGA-3ʹ) and respective controls (sh-NC, sh-con, mimic NC and anti-miR-NC) were synthesized by Ribobio Co., Ltd. (Guangzhou, China). pcDNA 3.1 vector (pcDNA-NC) and full-length SLC1A5 were used to generate SLC1A5 overexpression plasmid (pcDNA-SLC1A5). Cell transfection was executed using FuGENE6 (Roche) following the user’s manual.