Age-dependent upregulation of methylmalonic acid in serum promotes the progression of colorectal cancer, finds a study published in Cancer Cell International.
Human CRC cells HCT116 and SW480 were purchased from Cell Bank of Chinese Academy of Sciences in Shanghai. HCT116 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin . SW480 cells were cultured in L15 medium supplemented with 10% FBS and 1% penicillin–streptomycin.
For EDU assay, cells were treated with 50 μM EDU medium at 37 ℃ for 2 h, then fixed with 4% paraformaldehyde for 30 min. After permeabilized with 0.5% Triton-X, the cells were successively reacted with 1 × Apollo dyeing reaction solution and Hoechst 33,342 for 30 min. Finally, images were visualized using fluorescence microscope and analyzed by Image J and GraphPad Prism 6 software.
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