Apoptosis induced by ceramide in hepatocellular carcinoma Bel7402 cells.
OBJECTIVE
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.


CONCLUSIONS
Ceramide induces apoptosis in Bel7402 cells, related to Bcl-2 down-regulation.
Zhu
X F
XF
Department of Therapeutic Basis, Cancer Center, Sun Yat-sen University of Medical Sciences, Guangzhou 510060, China. xfzhu@gzsums.edu.cn
Zhang
X S
XS
Li
Z M
ZM
Yao
Y Q
YQ
Xie
B F
BF
Liu
Z C
ZC
Zeng
Y X
YX
D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D002518:Ceramides / Q000494:pharmacology*; D015536:Down-Regulation; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; D002518:Ceramides; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein