Knockdown of METTL3 in Cancer Cell Lines

Human gastric cancer cell TMK1, pancreatic cancer cell PANC-1, liver cancer cell Huh-7 were grown in DMEM media (Gibco) with 10% FBS (Gibco) and 1% 100X Pen/Strep (Gibco). Human colorectal cancer cell line HCT116, and esophageal cancer cell TE-1 were grown in IMEM and RPMI1640 media, respectively, with 10% FBS and 1% 100X Pen/Strep. Control siRNA and METTL3 siRNA were purchased from GenePharma (Shanghai, China) with reported sequences12 (link). Transfection was achieved by using Lipofectamine Lipofectamine 3000 (Invitrogen) with 20 nM of siRNAs. At 48 h post-transfection, the mRNA levels of METTL3, AKT1, mTOR, PIK3CA, and PTEN were checked by Q-PCR.