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Unless indicated otherwise, cystine-free medium was prepared by supplementing cystine-, methionine- and glutamine-free DMEM (Gibco, CA, USA) with 100 M methionine, 4 mM glutamine, 1 mM sodium pyruvate, 0.2% bovine serum albumin (BSA), 100 U/ml penicillin and 100 g/ml streptomycin

Further experiments demonstrated that the transfection of these cancer cells with ESM1 mimic upregulated the expression of MMP-2, MMP-3, MMP-9, VEGF, cyclooxygenase-2 and HIF-1 through the PI3K/AKT/mammalian target of rapamycin (mTOR) signaling pathway (53)

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10.1002/adfm.201901417 81 ZhangK.MengX.YangZ.DongH.ZhangX

Luo JL, Kamata H, Karin M (2005) IKK/NF-kappaB signaling: balancing life and deatha new approach to cancer therapy