The Effects of 5-Aza-2`-Deoxycytidine on DLC-1 Gene Expression, Methylation Level and Expression of Downstream Signaling Molecules Cdc42 in Multiple Myeloma

Author(s): Xianqi Feng, Ling Zhang, Shumin Nie, Zhan Su, Xue Shi, Yan Gao, Xiangyun Chen, Wenyuan Niu, Zhongguang Cui, Hongguo Zhao, Fanjun Meng, Chunting Zhao

Journal Name: Current Signal Transduction Therapy

Volume 10 , Issue 1 , 2015

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Objective: To investigate the effects of the DNA methyltransferase inhibitor 5-Aza-2'- Deoxycytidine (5-Aza-Dc) on DLC-1 gene expression, methylation level, and the expression of downstream signaling molecules Cdc42 in the human multiple myeloma cell line RPMI8226 cells.

Methods: The RPMI8226 cells were treated with 5-Aza-Dc, the methylation status of CpG island of DLC-1 gene was detected by bisulfate sequencing PCR(BSP) in RPMI8226 cells. The expression of DLC-1 and Cdc42 mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR). Enzyme-linked immunoabsorbent assay (ELISA) was used to detect the expression of Cdc42 protein.

Results: The methylation of DLC-1 gene was detected in the RPMI8226 cells without 5-Aza-Dc pretreament. DLC-1 gene methylation status was decreased after the 5-Aza-Dc treatment for 72h, and DLC-1 gene didn't display DNA methylation on the highest concentration of 5-Aza-Dc. DLC-1 mRNA was weakly expressed in the control group, and the expression was gradually increased in 5-Aza-Dc treatment group. The Cdc42 mRNA and protein expression of experimental group were significantly decreased and were dose-dependent compared with the control group (P <0.05).

Conclusion: The results of this research indicated that 5-Aza-Dc can effectively inhibit the methylation status of DLC-1 gene, reversal DLC-1 gene expression, and significantly decrease the expression of downstream signaling molecules Cdc42 mRNA and protein in RPMI-8226 cell.

Keywords: 5-aza-2`-deoxycytidine, Cdc42, DLC-1, methylation, multiple myeloma.

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Article Details

Year: 2015
Page: [31 - 35]
Pages: 5
DOI: 10.2174/1574362410666150311000910

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PDF: 96