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2. Regulation of Fas gene expression in HeLa cells as determined by modified RT-PCR

Author

Yoshizumi Shintani, Satoshi Kimura, Takeya Tsutsumi, Kazuhiko Koike, Hajime Fujie, Hiroshi Yotsuyanagi, and Kyoji Moriya

Subjects
Apoptosis, Polymerase Chain Reaction, Antibodies, HeLa, Cellular and Molecular Neuroscience, Humans, RNA, Messenger, fas Receptor, Molecular Biology, Pharmacology, Regulation of gene expression, Messenger RNA, biology, Cell Cycle, RNA, Cell Biology, Cell cycle, biology.organism_classification, Fas receptor, Molecular biology, Culture Media, Cell biology, Gene Expression Regulation, Neoplastic, Real-time polymerase chain reaction, Molecular Medicine, HeLa Cells
Abstract

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.

Published
1998

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