Xuefeng Liu1, Xianwei Dai, Bin Wu. 1. Department of Urology, Shengjing Hospital of China Medical University, Shenyang, China. zhaoyy@cmu2h.com
Abstract
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 micromol/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. (c) 2009 S. Karger AG, Basel.
OBJECTIVES: To investigate the effect of 5-Aza-CdR on methylation and expression of RASSF1A gene in the humanbladder cancer BIU87 cell line. METHODS: Cultured BIU87 cells were treated with increasing concentrations (0.1, 0.5, 1.0, 5.0 micromol/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. (c) 2009 S. Karger AG, Basel.
Authors: Surbhi Jain; Lijia Xie; Batbold Boldbaatar; Selena Y Lin; James P Hamilton; Stephen J Meltzer; Shun-Hua Chen; Chi-Tan Hu; Timothy M Block; Wei Song; Ying-Hsiu Su Journal: Hepatol Res Date: 2014-12-09 Impact factor: 4.288