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. 2019 Aug 14;9(21):6175–6190. doi: 10.7150/thno.35572

Figure 4.

Figure 4

Reducing the methylation levels of the QPCT promoter region by decitabine in RCC cells could increase the expression of QPCT and NF-κB (p65) bound to the QPCT promoter region, positively regulating its expression. (A) Sequenom MassARRAY Methylation of the QPCT promoter region in KETR-3 and OS-RC-2 cells after decitabine (0.5 μM) treatment for 48 h and control KETR-3 and OS-RC-2 cells (n=5). (B) CpG sites that had differences between the two groups in the QPCT promoter region (n=5). (C) QPCT mRNA expression in KETR-3 and OS-RC-2 cells after decitabine (0.5 μM) treatment and control KETR-3 and OS-RC-2 cells (n=3). (D) QPCT protein in KETR-3 and OS-RC-2 cells after decitabine (0.5 μM) treatment and control KETR-3 and OS-RC-2 cells (n=3). (E) qPCR analysis of NF-κB (p65) mRNA in 16 pairs of sunitinib-responsive and nonresponsive RCC tissues. (F) Western blot analysis of NF-κB (p65) protein in 15 pairs of sunitinib-responsive and nonresponsive RCC tissues. (G) ChIP analysis demonstrated that NF-κB (p65) binds to the QPCT promoter region and increased NF-κB (p65) binding to the QPCT promoter after inhibiting the methylation levels of QPCT. (H) QPCT mRNA (above) and protein (below) expression in ACHN and OS-RC-2 cells after triptolide (10 nM) treatment for 72 h and control ACHN and OS-RC-2 cells (n=3). (I) QPCT mRNA (above) and protein (below) expression in 786-O and KETR-3 cells after BetA (5 μM) treatment for 72 h and control 786-O and KETR-3 cells (n=3). Results are presented as the means ± SD. *p<0.05, **p<0.01.