Kato 3

Five human GC cell lines, SNU-5, AGS, MKN-74, HGC-27, and KATO III and the normal gastric mucosal epithelial cell line GES-1 were purchased from Wuhan Procell Life Science & Technology (Wuhan, China). These cells were recovered and cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Gibco, Grand Island, NY) supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (100 mg/mL) under saturated humidity, 5% CO₂, and 37°C. When cell confluence reached 90%, serial passaging was performed. The cells were detached with 0.25% trypsin until the cells become round and gaps appeared, which was subsequently terminated by addition of FBS culture solution. Next, the cells were pipetted into a single-cell suspension. Quantitative real-time polymerase-chain reaction (qRT-PCR) was then employed to determine miR-642b-3p expression in GC cell lines [22 (link)].

GC cells (GES-1, AGS, HGC-27, KATO III, MKN-1, MKN-45) were purchased from Procell Life Science & Technology (Wuhan, China), and the cells were cultured according to the manufacturer’s instructions. The cell lines were cultured in RPMI-1640 medium (Gibco, USA) and were supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin/streptomycin (Gibco).

The Ethics Committee of Hubei University of Medicine (Shiyan, China) approved all research. Archived GC tissue specimens were collected from patients undergoing surgery in Taihe Hospital without radio- or chemotherapy after obtaining written informed consent. The samples were divided into two parts, one was fixed in formalin and another was stored at -80 °C. GC cell lines (KATO-III, BGC-823, MGC-803, SGC-7901, MKN-74, HGC-27) and gastric mucosal epithelial cell line (GES-1) were purchased from Procell (Wuhan, China). Cells were cultured in RPMI-1640 or Hams F12 medium (Gibco, USA) containing 10% fetal bovine serum (FBS; Gibco).

Normal gastric mucosal epithelial cells (GES-1) and GCcells (AGS, HGC-27, KATO3, MKN-45) were purchased from Procell Life Science & Technology (Wuhan, China), and the cells were cultured according to the manual instructions. A lactate dehydrogenase A (LDHA) inhibitor (GSK2837808A, MCE) was used to treat HGC cell lines. The HGC cell lines were cultured in six-well plates, and plasmids were transfected using Lipofectamine 3000 according to the instructions. The target sequences of the short hairpin RNA (shRNA) were as follows: PLOD2-RNAi (8407-1), caGCAAGTGTCCTTAAGTCAA; PLOD2-RNAi (8408-1), ggAAATGGACCCACCAAGATT; PLOD2-RNAi (8409-1), ctTTGCCGAAATGCTAGAGAA.