ABSTRACT
Objective: To investigate the effect of sodium butytate (different concentrations) on the growth and proliferation of rat liver oval cell line WBF344, and to discuss the conditions and tules for sodium butyrate-inducd WB-F344 cells differentiation into biliary lineage in vitro. Methods: WB-F344 cells were treated with sodium butyrate (0.70, 2.25, 3.75, 4.5 mmol/L) and the cell growth and morphological changes were observed; routinely cultured WB-F344 cells were taken as control. The changes of CK19 protein expression were examined immunohistochemically after WB-F244 cells were treated with 3.75% sodium butyrate; and the expression of phenotypic markers, such as γ-glutamyltransferase (GGT), β4-integrin, CK19, AFP and ALB at toRNA level were determined by RT-PCR. Untreated WB-F344 cells were used as blank control. Results: We found that sodium butyrate inhibited the growth of WB-F344 cells. The optical densities were significantly decreased in 3.75 and 4.5 mmol/L groups compared with that in control group(P<0.01); but no significant difference was found between 0.75, 2.25 mmol/L groups with control group. WB-F344 cells treated with 3.75, 4.5 mmol/L sodium butyrate became larger and round, with increased nuclei and decreased nucleus to cytoplasm ratio; those treated with 0.75, 2.25 mmol/L had no obvious changes. Immunohistochemical results showed that sodium butyrate significantly increased CK19 expression compared with control group ([92.3±1.1]% vs [1.3±0.2]%, P<0.01). RT-PCR showed increased expression of β4-integrin in sodium butyrate treated groups, but not in control group; the expression of GGT and CK19 was higher than that of control group. Alpha-fetoprotein (AFP) expression was observed in blank control group, but not in sodium butyrate treated cells. Albumin expression was not detected in the 2 groups. Conclusion: Sodium butyrate at 3.75 mmol/L is suitable for inducing WB-F344 cells differentiate into the biliary lineage in vitro.
ABSTRACT
Objective:Establishing the hepatic stem cell(WB-F344)line that expresses human interleukin-2 stably and effectively.Methods: We constructed the retroviral vector Plpcx-il-2 containing human interleukin-2 gene, and transfected it into PT67 packaging cells.The infectious viruses were used to infect the hepatic stem cell WB-F344,then the puromycin resistant clones were acquired after puromycin selection.The expression of IL-2 was detected using RT-PCR,immunocytochemistry, Western blot and indirect immunofluorescence, respectively.Results: The retroviral vector containing human IL-2 gene was constructed.The titer of virus in the supernatant reached 10~5 CFU/ml after transfecting PT67 packeting cells.The puromycin resistant clones were acquired after infection of WB-F344 and the cell line WB-F344/Plpcx-IL-2 expressing human interleukin-2 stably was established.Conclusion: By retroviral-mediated transfecting human interleukin-2 gene, the hepatic stem cell stably and effectively expressing interleukin-2 are obtained.This provides a good basis for further research on immuno-gene therapy of liver cancer.
ABSTRACT
Objective:To investigate the effect of sodium butytate (different concentrations) on the growth and proliferation of rat liver oval cell line WB-F344, and to discuss the conditions and rules for sodium butyrate-inducd WB-F344 cells differentiation into biliary lineage in vitro. Methods: WB-F344 cells were treated with sodium butyrate (0.75, 2.25, 3.75, 4.5 mmol/L) and the cell growth and morphological changes were observed; routinely cultured WB-F344 cells were taken as control. The changes of CK19 protein expression were examined immunohistochemically after WB-F244 cells were treated with 3.75% sodium butyrate; and the expression of phenotypic markers, such as ?-glutamyltransferase (GGT) ,?4-integrin, CK19, AFP and ALB at mRNA level were determined by RT-PCR. Untreated WB-F344 cells were used as blank control. Results: We found that sodium butyrate inhibited the growth of WB-F344 cells. The optical densities were significantly decreased in 3.75 and 4.5 mmol/L groups compared with that in control group(P