Research Paper Volume 13, Issue 3 pp 4045—4062

ALKBH5-mediated m6A demethylation of FOXM1 mRNA promotes progression of uveal melanoma

High ALKBH5 expression is associated with worse prognosis in patients with UM and promotes UM growth in vivo (A) Survival curves of UM patients expressing ALKBH5 at high and low levels in the TCGA cohort from the online GEPIA database (P = 0.0023). (B) Expression of ALKBH5 protein (upper panel) and mRNA (bottom panel) levels in UM cells was detected by western blot and qRT-PCR. (C) ALKBH5 knockdown efficiency was verified at the protein and mRNA levels in C918 (left panel) and MuM-2B (right panel) by western blot and qRT-PCR. (D) Knockdown of ALKBH5 effectively inhibited UM subcutaneous tumor growth in nude mice (n=5). (E) The growth curves of C918 stably transduced with shALKBH5 in nude mice were significantly dampened compared with those of C918 cells transduced with control plasmid. (F) Histogram shows the mean tumor weights from the shALKBH5 and control groups. (G) Sections of tumors were stained with anti-ALKBH5, anti-Ki67, and anti-cleaved-Caspase 3 antibodies by IHC staining. Mean ± SEM, t-test, *P P P

Figure 1. High ALKBH5 expression is associated with worse prognosis in patients with UM and promotes UM growth in vivo (A) Survival curves of UM patients expressing ALKBH5 at high and low levels in the TCGA cohort from the online GEPIA database (P = 0.0023). (B) Expression of ALKBH5 protein (upper panel) and mRNA (bottom panel) levels in UM cells was detected by western blot and qRT-PCR. (C) ALKBH5 knockdown efficiency was verified at the protein and mRNA levels in C918 (left panel) and MuM-2B (right panel) by western blot and qRT-PCR. (D) Knockdown of ALKBH5 effectively inhibited UM subcutaneous tumor growth in nude mice (n=5). (E) The growth curves of C918 stably transduced with shALKBH5 in nude mice were significantly dampened compared with those of C918 cells transduced with control plasmid. (F) Histogram shows the mean tumor weights from the shALKBH5 and control groups. (G) Sections of tumors were stained with anti-ALKBH5, anti-Ki67, and anti-cleaved-Caspase 3 antibodies by IHC staining. Mean ± SEM, t-test, *P < 0.05, **P < 0.01, ***P < 0.001.