TY - JOUR
T1 - Butyrate synchronization of hepatocytes
T2 - Modulation of cycling and cell cycle regulated gene expression
AU - Gupta, Sanjeev
AU - Alpini, Gianfranco
AU - Wmuru, Ravikumar P.
AU - Hurston, Ethel
AU - Shafritz, David A.
N1 - Funding Information:
Supported in part by Public Health Service grants from the NIH (DK-01909, DK-17609, P30-41296 to the Marion Bessin Liver Research Center and P30 CA 13330 to the Cancer Research Center at Einstein). This work was presented in part at the Annual Meeting of the American Association for Studies on Liver Diseases, Chicago, October 1989 and published as a n abstract (Hepatology 1989; 10: 622). S. G. is the recipient of a Clinical Investigator Award from the NIDDK.
PY - 1994
Y1 - 1994
N2 - To develop a model for studies of liver growth control, we characterized cell cycle synchronization of liver-derived cells with sodium butyrate. Exposure of cultured HTC (rat hepatoma) cells to 5 mM butyrate arrested cell growth in a reversible manner. Flow cytometric analysis revealed that butyrate-treated HTC cells were restricted in G0/G1, as well as S/G2M phases. After release from butyrate arrest, HTC cells underwent synchronous cycles of DNA synthesis and transited through S phase. Inhibition of cell growth by butyrate was associated with a complex pattern of cell cycle regulated gene expression, including a decoupling of c-fos and c-jun gene expression. Transcription of c-fos, as well as c-jun increased with butyrate arrest, whereas steady rate mRNA levels of c-jun only were increased, suggesting additional regulation of c-fos. In addition, butyrate-arrested cells exhibited a transcriptionally determined accumulation of H3 histone, C-Ha-ras and ornithine decarboxylase mRNAs, suggesting that cell cycle-related check points following the onset of S phase were modulated. An increase in c-myc mRNA levels in butyrate-arrested cells was post-transcriptionally regulated. After release from butyrate-arrest, the abundance of immediate early, as well as S phase regulated, gene expression changed coordinately with S phase cell transitions. Thus, exposure of HTC cells to butyrate modulates cell cycle regulated gene expression, inhibits cycling, and results in accumulation of cells in specific compartments. Synchronization of liver cells with butyrate should, therefore, provide a useful model for defining cell cycle-related events in response to various mitogenic stimuli.
AB - To develop a model for studies of liver growth control, we characterized cell cycle synchronization of liver-derived cells with sodium butyrate. Exposure of cultured HTC (rat hepatoma) cells to 5 mM butyrate arrested cell growth in a reversible manner. Flow cytometric analysis revealed that butyrate-treated HTC cells were restricted in G0/G1, as well as S/G2M phases. After release from butyrate arrest, HTC cells underwent synchronous cycles of DNA synthesis and transited through S phase. Inhibition of cell growth by butyrate was associated with a complex pattern of cell cycle regulated gene expression, including a decoupling of c-fos and c-jun gene expression. Transcription of c-fos, as well as c-jun increased with butyrate arrest, whereas steady rate mRNA levels of c-jun only were increased, suggesting additional regulation of c-fos. In addition, butyrate-arrested cells exhibited a transcriptionally determined accumulation of H3 histone, C-Ha-ras and ornithine decarboxylase mRNAs, suggesting that cell cycle-related check points following the onset of S phase were modulated. An increase in c-myc mRNA levels in butyrate-arrested cells was post-transcriptionally regulated. After release from butyrate-arrest, the abundance of immediate early, as well as S phase regulated, gene expression changed coordinately with S phase cell transitions. Thus, exposure of HTC cells to butyrate modulates cell cycle regulated gene expression, inhibits cycling, and results in accumulation of cells in specific compartments. Synchronization of liver cells with butyrate should, therefore, provide a useful model for defining cell cycle-related events in response to various mitogenic stimuli.
KW - Butyrate
KW - Cell cycle
KW - Gene expression
KW - Hepatocyte
KW - Protooncogenes
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U2 - 10.3109/08977199409000235
DO - 10.3109/08977199409000235
M3 - Article
C2 - 7946406
AN - SCOPUS:0028229286
SN - 0897-7194
VL - 10
SP - 171
EP - 180
JO - Growth Factors
JF - Growth Factors
IS - 3
ER -