Mkn 74

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)].

Human GC cell lines (AGS and MKN-74) and Mouse Forestomach Carcinoma (MFC) cell line were purchased from the Procell Life Science and Technology (Wuhan, China). These cells were cultured in RPMI 1640 medium (Procell Life Science and Technology, Wuhan, China) with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA). Culture medium was changed every 2 days. Cells were pretreated with SR1078 (RORα activator, GC16392, GlpBio, Montclair, CA, USA), 3PO (3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one, PFKFB3 inhibitor, MCE, USA), DHEA (Dehydroepiandrosterone, G6PD inhibitor, MCE, USA) and glucose for 24 h. Cells were pretreated with TGF-β1 (Transforming growth factor beta 1, Cayman chemical, USA) for 48 h. SR1078 was dissolved in phosphate buffer saline (PBS). 3PO, DHEA, TGF-β1 and fluorouracil (GC14466, GlpBio, Montclair, CA, USA) were dissolved in dimethyl sulfoxide (DMSO).

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).