Effects Of 5-aza-CdR On Proliferation And Invasion In MDA-MB-231 Cell

Objective:To investigate the effects of DNA methylation inhibitor 5-Aza-2’-deoxyeytidine(5-Aza-Cd R) with the proliferation and invasion of human breast cancer MDA-MB-231 cell line and detect expression level of genes with aberrant methylation such as WIF1. It will built a solid foundation for new therapeutic targets and chemotherapy drugs of breast cancer.Methods:1. Cell culture and drug treatment: The MDA-MB-231 cells were cultured with RPMI 1640 medium. After being subcultured, the human breast cancer MDA-MB-231 cells were treated with RPMI 1640 culture solution containing different concentrations( 1μmol/L、5μmol/L、 10μmol/L、20μmol/L) of 5-Aza-Cd R. The MDA-MB-231 cells without the intervention of 5-Aza-Cd R were used as the control group. 2. The proliferation of MDA-MB-231 cells was detected by using a MTT cell proliferation assay. 3.The cell scratch test was carried out to observe the invasion distance of the cell. 4. The m RNA and protein expression of E-cadherin,WIF1, β-catenin in breast cancer MDA-MB-231 cells was showed by using a real-time reverse transcription polymerase chain reaction(RT-PCR) assay and Western Blot.Results:1.The proliferation of human breast caner MDA-MB-231 cells were significantly inhibited in a dose-dependent and a time-dependent manner.2.The invasion of MDA-MB-231 cells was quantified by cell scratch test: After 24 hours,the healing rate of MDA-MB-231 cells of 5μmol/L、10μmol/L group was(30.0%±2.0%)、(24.6%±2.4%) lower than control group45.0%±1.3%). 48 hours later, the difference was also significant, The healing rate of MDA-MB-231 cells of 5μmol/L、10μmol/L group was(37.3%±2.2%)、(25.7%±2.4%)control was(55.3%±2.2%)(P<0.05).It is statistically significant;3. RT-PCR and Western blot indicated that the expression of E-cadherin、WIF1 significantly increase in MDA-MB-231 cells treated with 10μmol/L 5-Aza-Cd R.It has a time-dependent,while β-catenin decreased with time goes;4..the expression of β-catenin markedly decrease in MDA-MB-231 cells treated with 10μmol/L 5-Aza-Cd R and have a time-dependent manner.Conclusion:5-Aza-Cd R could inhibit proliferation and invasion ability of MDA-MB-231 cells and its mechanism related to increasing the expression of WIF1 and E-cadherin and decreasing β-catenin in MDA-MB-231 cells treated with 5-Aza-Cd R.