Apoptosis induced by ceramide in hepatocellular carcinoma Bel7402 cells
Abstract
"AIM:
To study the biological function of ceramide signaling in Bel7402 cells.
METHODS:
Inhibition of cell growth was assayed using MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was detected by electrophoresis and flow cytometry. The levels of protein p53, Bcl-2, and Bax were measured with Western blot.
RESULTS:
Bel7402 cells treated with C2-ceramide underwent cell proliferation inhibition. IC50 value was 14.28 mumol.L-1. After treatment of Bel7402 with ceramide, the morphologic changes including reduction in volume, nuclear chromatin condensation, fluorescence strength were observed. SubG1 peaks were detected on flow cytometry (FCM). Agarose gel electrophoresis of DNA from cells treated with ceramide revealed ""ladder"" pattern. The Western blot assay from cell extracts showed that the levels of protein p53 were decreased after ceramide treatment. The levels of protein Bcl-2 were decreased also. But the levels of Bax protein showed no difference between untreated cells and treated cells.
CONCLUSION:
Ceramide induces apoptosis in Bel7402 cells, related to Bcl-2 down-regulation."
Keywords:
To study the biological function of ceramide signaling in Bel7402 cells.
METHODS:
Inhibition of cell growth was assayed using MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was detected by electrophoresis and flow cytometry. The levels of protein p53, Bcl-2, and Bax were measured with Western blot.
RESULTS:
Bel7402 cells treated with C2-ceramide underwent cell proliferation inhibition. IC50 value was 14.28 mumol.L-1. After treatment of Bel7402 with ceramide, the morphologic changes including reduction in volume, nuclear chromatin condensation, fluorescence strength were observed. SubG1 peaks were detected on flow cytometry (FCM). Agarose gel electrophoresis of DNA from cells treated with ceramide revealed ""ladder"" pattern. The Western blot assay from cell extracts showed that the levels of protein p53 were decreased after ceramide treatment. The levels of protein Bcl-2 were decreased also. But the levels of Bax protein showed no difference between untreated cells and treated cells.
CONCLUSION:
Ceramide induces apoptosis in Bel7402 cells, related to Bcl-2 down-regulation."