Figure 1.
Determination of resveratrol sensitivities of LN-18 and PBC cells to resveratrol.
(A) H&E morphological staining performed on UW228-3, LN-18 and PBC cells without (normal culture) and with 100 µM resveratrol treatment for 48 hours. The results of synaptophysin-oriented ICC and TUNEL assay are shown in the upper and the lower insets for each of the main images, respectively. (B) Flow cytometry fractionation of cell cycles and apoptotic cells in UW228-3, LN-18 and PBC cell populations without (NC) and with (Res) 100 µM resveratrol treatment for 48 hours. *, indicates the peak of apoptotic cells.
Figure 2.
The metabolic patterns analyses of resveratrol in LN-18 and PBC cells.
(A) The MS/MS spectrum of M1, M2 and M3 correspond to 1, 2 and 3 of the HPLC peaks. 4, an unidentified compound produced predominantly in PBCs. (B) Shimadzu LCMS-IT-TOF-based HRMS analysis of resveratrol metabolites in LN-18 cells and PBCs. Arrows labeled as trans-resveratrol, cis-resveratrol, resveratrol monosulfate and resveratrol glucuronide? indicate the exact [M–H]- molecular ion weight of 227.0698 (C14H11O3, calculated m/z 227.0708), 227.0697 (C14H11O3, calculated m/z 227.0708), 307.1105 (C14H11SO6, calculated m/z 307.0276) and 403.0992 (C20H19O9, calculated m/z 403.1035), respectively.
Figure 3.
Resveratrol upregulated SULT1A1, 1C2 and 4A1 expression.
(A) ICC evaluation of SULT1A1, 1C2 and 4A1 expression in PBC, UW228-3 and LN-18 cells without (N) and with (R) resveratrol treatment. (B) Western blot analyses of SULT1A1, 1C2 and 4A1 expression in LN-18 cells without (N) and with (R) resveratrol treatment and compared with that in normal control PBCs and resveratrol-sensitive control medulloblastoma UW228-3 cells. β-actin was used as loading control and for calculation of SULT expression levels/densitometry scan of Western blotting results. *Compared with normal cultured LN-18 and UW228-3 cells, respectively; *represents statistical significance (p<0.05).
Figure 4.
The expression of SULT1A1, 1C2 and 4A1 in glioblastomas tissues.
Immunohistochemical illustration of differential expression patterns of SULT1A1, 1C2 and 4A1 in two cases of glioblastomas (GM). Medulloblastoma (MB) and its surrounding noncancerous cerebellum tissue (NC) were cited as controls.
Table 1.
Immunohistochemical profiling of SULT expression in human brain tissues.
Figure 5.
Evaluation of STAT3 and PIAS3 statuses in UW22-3, LN-18 and PBC cells.
STAT3-oriented RT-PCR (A), immunocytochemical and immunofluorescent illustration of intracellular distribution of STAT3 (B) and its inhibitor PIAS3 (C) in LN-18 cells without (N) and with (R) 100 µM resveratrol treatment. Immunofluorescence results were shown in the insets for Fig 5B and 5C. Corresponding data obtained from PBC and UW228-3 cells were used as controls.
Figure 6.
Sensitivities of LN-18 and PBC cells to STAT3 inhibitor AG490.
(A) Immunofluorescence illustration of intracellular distribution of STAT3 in LN-18 and PBC cells without (N) and with (AG490) 60 µM AG490 treatment. Arrows indicate the cells shown in the insets in high magnification (X400). (B) Flow cytometry analysis revealed reduction of G1-phase cells, accumulation of S-phase cells and induction of apoptosis (blue peak) in AG490-treated LN-18 cell population. The cell cycle progression was almost unchanged in PBC cells. *, indicates the peak of apoptotic cells. (C) H&E morphologic examination of LN-18 cells under normal culture or incubated with 100 µM trans-resveratrol, 60 µM AG490 or resveratrol/AG490 mixture for 48 hours (Main images). Small images: immunocytochemical illustration of SULT1A1, 1C2 and 4A1 expression in LN-18 cells treated by 60 µM AG490 without/with 100 µM resveratrol supplementation. Normally cultured and 100 µM resveratrol-treated LN-18 cells were cited as controls.