Regulation of Fas gene expression in HeLa cells as determined by modified RT-PCR.
We determined human Fas messenger RNA (mRNA) levels in HeLa cells using a 'mutagenic' reverse transcription-polymerase chain reaction, which quantitates mRNA levels using the corresponding genomic DNA as an internal control. The expression level of Fas mRNA was very low in serum-deprived quiescent HeLa cells. In conjunction with the start of cell-cycle progression upon the addition of serum to culture medium, the Fas mRNA level gradually increased, reached its peak at 36 h and returned to the basal level after 48 h. HeLa cells at 36 h exhibiting a high level of Fas mRNA expression were more susceptible to the anti-Fas antibody apoptotic signal. Thus, the regulation of Fas expression is associated with cell-cycle progression, and this method for Fas mRNA detection may be useful, particularly for the analysis of small amounts of samples.
Yotsuyanagi
H
H
First Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Shintani
Y
Y
Moriya
K
K
Fujie
H
H
Tsutsumi
T
T
Kimura
S
S
Koike
K
K
D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D017209:Apoptosis / Q000276:immunology / Q000502:physiology; D002453:Cell Cycle / Q000502:physiology; D003470:Culture Media / Q000737:chemistry; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D006367:HeLa Cells / Q000166:cytology; D006801:Humans; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D019014:fas Receptor / Q000235:genetics* / Q000276:immunology
D000906:Antibodies; D003470:Culture Media; D012333:RNA, Messenger; D019014:fas Receptor