Fig 1.
LPCAT1 knockdown alters CRPC progression.
(A): LPCAT1 Si-RNA was transfected into C4-2 and PC-3 cells, and transfection efficiency was demonstrated. (B): The ability of cell proliferation was measured at 0, 24, 48 and 72 h after transfection. (C) and (D): The ability of cell migration and invasion were determined at 0, 24 and 48h after transfection. (E): The PAF concentration in cell cultures of Si-LPCAT1 group and control at 48 h after transfection. (F): Cell proliferation assay were measured at 48 h after C4-2 and PC-3 cells in Si-LPCAT1 group were treated with increasing concentration of PAF (10−8–10−6 M) and the control group. (G) and (H): Cell migration and invasion assay were measured at 48 h after increasing concentration of PAF (10−8–10−6 M) in Si-LPCAT1 group and control group, and ABT-491 (10−5 M) was used to pretreat cells induced by PAF (10−6 M). Values are presented as mean percent control ± s.e.m or mean ± s.e.m. * P < 0.05 and ** P < 0.01 compared with control; # P < 0.05 and ## P < 0.01 compared with Si-LPCAT1 treated with PAF (0 M); & P < 0.05 and && P < 0.01 compared with Si-LPCAT1 treated with PAF (10−6 M).
Fig 2.
LPCAT1 overexpression alters motility in CRPC cells.
(A): LPCAT1 vector and empty vector (control) was transfected into C4-2 cells, and the transfection efficiency was demonstrated. (B): the ability of cell migration and invasion was determined at 0, 24 and 48 h after transfection. (C): The PAF concentration in cell cultures of LPCAT1 group and control group. (D) and (E): Cell migration and invasion assay were measured at 48 h after increasing concentration of PAF-AH (1–50 μg/ml) and ABT-491(10−7–10−5 M) in LPCAT1 group and control group. Values are presented as mean percent control ± s.e.m or mean ± s.e.m. * P < 0.05 and ** P < 0.01 compared with control.
Fig 3.
LPCAT1 overexpression alters CRPC cell growth.
(A): The ability of Cell proliferation was measured at 0, 24, 48 and 72 h after transfection. (B): Colony formation efficiency was measured in LPCAT1 group and control group. (C): cell cycle was measured in LPCAT1 group and control group. (D): Tumorigenicity assay in 6 nude mice was performed and tumors weights in LPCAT1 and control groups were measured. (E): The PAF concentration was measured at 48 h after PAF-AH treatment. (F): The ability of cell proliferation in LCPAT1 group with different concentration PAF-AH treatment and control group was measured at 0, 24, 48 and 72h. Values are presented as mean percent control ± s.e.m or mean ± s.e.m. * P < 0.05 and ** P < 0.01 compared with control.
Fig 4.
Androgen promotes LPCAT1 nuclear relocalization.
(A): Intra-nuclear LPCAT1 expression in LPCAT1 and control groups and cells were cultured with ordinary FBS containing a certain amount of androgen. (B): The effect of the increased concentrations of DHT on intra-nuclear LPCAT1 expression of C4-2 cells cultured in Dextran Stripped FBS was detected. (C): The effect of DHT (10−6 M), DHT (10−6 M) +Flu and DHT (0 M) on the proliferation ability of C4-2 cells cultured in Dextran Stripped FBS was detected. (D): Tumors weights in 6 nude mice with LPCAT1 and control groups after castration. (E): Activated RNA polymerase (RNAP) Ⅱ expression (up) and total RNA synthesis (down) were measured in LPCAT1 and control groups cells with ordinary FBS culture. (F): Histone H4 S47A mutation was designed and transfected into LPCAT1 and control groups, and activated RNA polymerase (RNAP) Ⅱ expression (up) and total RNA synthesis (down) were measured in WT and mutation groups. Values are presented as mean percent control ± s.e.m or mean ± s.e.m. * P < 0.05 and ** P < 0.01 compared with control. # P < 0.05 and ## P < 0.01 compared with LPCAT1 overexpressing cells in WT group.
Fig 5.
LPCAT1 expression regulates paclitaxel sensitivity.
(A): Cell viability was determined in LPCAT1 and control groups with the increasing concentration of paclitaxel treatment (1, 10 and 100 nM). (B): Cell apoptosis in LPCAT1 and control groups with paclitaxel (10 nM) treatment. (C) and (D): Cells were injected into 6 nude mice to initiate tumorigenesis, with paclitaxel treatment administered 14 days post inoculation. Tumor volumes at different time points (C) and tumors weights (D) at excision. Values are presented as mean ± s.e.m. * P < 0.05 and ** P < 0.01 compared with control.