Fig 1.
Proliferation assay in BAP1 knockdown GBC cell line.
BAP1 expression of the GBC cell line G-415 was down-regulated by siRNA and the proliferative ability was measured by MTS assay. No significant difference was observed between each siRNA group and the control. The vertical axis shows the relative intensity compared to the intensity at day 1, and the horizontal axis shows the number of cell culture days.
Fig 2.
Migration assay in BAP1 knockdown GBC cell line.
(a) A diagram immediately after scratching in the confluently cultured cells (day 1) and the narrowed gap by migrating cells (day 2) are shown. (b) The relative increase of the migration area compared to the control group after 24 hours was significantly higher in the BAP1 knocked-down cells.
Fig 3.
Invasion assay in BAP1 knockdown GBC cell line.
(a) The GBC cell line G-415 was transfected with siRNA against BAP1 and was seeded on a Matrigel invasion chamber and the infiltrated cells on the lower surface of the chamber were stained. (b) Compared to the control group, the number of infiltrated cells tended to increase in both siRNA1 and siRNA2, and the number of infiltrated cells was significantly higher in the siRNA1.
Fig 4.
Drug sensitivity test in BAP1 knockdown GBC cell line.
Cell viability of BAP1 knocked-down GBC cell line G-415 under the agents of gemcitabine (GEM), cisplatin (CDDP), fluorouracil (5-FU), sodium valproate, 5-azacytidine, and bortezomib was measured by MTS assay. No change in sensitivity was observed in GEM, CDDP, 5-FU, sodium valproate or 5-azacytidine. Bortezomib showed a significant decrease in sensitivity in the siRNA1 and siRNA2 group compared to the control.
Table 1.
Clinical characteristics of the 47 cases of GBC patients.
Fig 5.
Kaplan–Meier survival analysis of BAP1 expression.
Kaplan–Meier survival curves for 41 GBC cases except for in-hospital deaths and deaths from other diseases show a significantly poorer prognosis in the BAP1 low expression group.
Fig 6.
BAP1 expression and detected mutations in clinical GBC (ID 12).
(a) Since the BAP1 staining concentration was low in the cancer part (Ca 40.4 < Liver 70.3), ID12 was categorized to the BAP1 low expression group. (b) The nonsense mutation on exon 8 (c.587G>A (p.Trp96Ter)) and the missense mutation on exon 4 (c.128T>A (p.Val43Glu), c.197T>A (p.Val66Glu)) were observed.
Table 2.
Summary of clinical characteristics, BAP1 expression, mutation, homozygous deletion and methylation in the GBC patients.
Table 3.
Summary of mutations and homozygous deletion in the 47 cases of GBC patients.