Objectives: To investigate the effect of 5-Aza-CdR on methylation and expression of RASSF1A gene in the human bladder cancer BIU87 cell line. Methods: Cultured BIU87 cells were treated with increasing concentrations (0.1, 0.5, 1.0, 5.0 μmol/l) of 5-Aza-CdR. MTT was used to detect the proliferation of BIU87 cells. The methylation status and expression level of RASSF1A gene in BIU87 cells were analyzed by methylation-specific polymerase chain reaction and RT-PCR before and after treatment with 5-Aza-CdR. Meanwhile, Western blot was used to detect the changes of RASSF1A protein. Results: In this study we found that the growth velocity of the BIU87 cell line was suppressed to various degrees after having been treated with different concentrations of 5-Aza-CdR, and the survival rate decreased with increasing 5- Aza-CdR concentrations. Besides, the methylation status of RASSF1A was obviously reversed and mRNA was re-expressed after treatment with 5-Aza-CdR. Expression of RASSF1A protein increased with increasing 5-Aza-CdR concentrations. Conclusions: Our results suggest that the tumor-suppressive effect of 5-Aza-CdR may result from reactivation of silenced RASSF1A through a demethylation function.

Keywords:
5-Aza-CdR, Bladder carcinoma, RASSF1A
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© 2009 S. Karger AG, Basel
2009
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