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Fig. 4 | Cancer Cell International

Fig. 4

From: MeCP2 confers 5-fluorouracil resistance in gastric cancer via upregulating the NOX4/PKM2 pathway

Fig. 4

MeCP2 promoted NOX4 transcription by binding to its promoter in 5-FU-resistant GC cells. A The MeCP2 binding site on the promoter region of NOX4. B–D ChIP RT-PCR of NOX4 was performed respectively with anti-MeCP2 antibody, anti-GFP antibody after transfection with GFP-MeCP2 plasmid, and anti-GFP antibody after transfection with Ctrl (GFP plasmid), WT (GFP-MeCP2 plasmid), MT1 (GFP- Mutation type 1) or MT2 (Mutation type 2). E BGC-823/5-FU cells were transfected with pGL3-luc, pGL3-NOX4-luc (target sequence of NOX4 promoter region), or pGL3-NOX4-luc + Methylation. Luciferase activity was determined at 48 h post-transfection. Renilla luciferase served as the internal control. Data are shown as mean ± SEM (*p < 0.001, as compared with pGL3-luc; #p < 0.001, as compared with pGL3-NOX4-luc; Student's t-test). F BGC-823/5-FU cells were transfected with NC-siRNA, MeCP2 siRNAs, MeCP2 overexpression plasmid, or null vector, and treated with pGL3-NOX4-luc or pGL3-NOX4-luc + Methylation, respectively. Luciferase activity was determined at 48 h post-transfection. Data are shown as mean ± SEM (*p < 0.001, as compared with NC-siRNA; #p < 0.001, as compared with null vector; Student's t-test). G MeCP2 and NOX4 levels were positively correlated in 81 pairs of GC tissue and normal para-carcinoma tissue (PCT). The 2−ΔΔCt values of MeCP2 and NOX4 mRNA were subjected to a Pearson correlation analysis (r = 0.466, n = 81, p < 0.0001, Pearson's correlation). H NOX4 mRNA expression in GC tissues and PCTs. Data are shown as mean ± SEM (*p < 0.01, Student's t-test). I NOX4 protein expression in GC tissues versus normal PCTs by immunohistochemistry. J NOX4 protein expression in GC tissues versus normal PCTs (pooled sample). K Silencing MeCP2 down-regulated the expression of NOX4 in 5-FU-resistant BGC-823 and SGC-7901 cells. L Overexpressed MeCP2 promoted the expression of NOX4 in the cells

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