5-aza-CdR对乳腺癌MDA-MB-231细胞生长增殖及迁移能力的影响

发布时间：2018-02-01 22:43

本文关键词： 5-aza-CdR 乳腺癌 WIF1 E-cadherin β-catenin 增殖迁移　出处：《南华大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的:研究5-氮杂-2'-脱氧胞苷对乳腺癌MDA-MB-231细胞系增殖及迁移的影响,为探索乳腺癌新的治疗靶点及化疗药物奠定基础。方法:1.细胞培养及药物处理:体外培养人乳腺癌细胞株MDA-MB-231,经传代后,分别用含有不同浓度(1μmol/L、5μmol/L、10μmol/L、20μmol/L)5-氮杂2-脱氧胞苷(5-Aza-Cd R)的RPMI1640培养液进行培养,并设不含该药物的对照组;2.用噻唑蓝MTT法测定不同浓度5-Aza-Cd R作用MDA-MB-231细胞株不同时间的OD值,检测细胞增殖速度的变化;3.采用划痕试验检测各组乳腺癌细胞株MDA-MB-231的迁移距离;4.采用逆转录PCR法(Reverse transcription-polymerase chain reaction)、Western Blot检测10μmol/L 5-Aza-Cd R作用不同时间后E-cadherin、WIF1及β-catenin m RNA和蛋白的影响;5.采用SPSS18.0分析各处理组与对照组细胞抑制率、相关基因m RNA及蛋白表达水平的差异。结果:1.MTT结果显示,5-Aza-Cd R对MDA-MB-231细胞生长有明显抑制作用,并且存在剂量及浓度依赖性反应关系;2.细胞迁移显示:划痕24小时后,5μmol/L、10μmol/L药物处理组MDA-MB-231细胞愈合率分别为(30.0%±2.0%)、(24.6%±2.4%),较对照组(45.0%±1.3%)低,48小时后,5μmol/L、10μmol/L药物处理组MDA-MB-231细胞愈合率分别为(37.3%±2.2%)、(25.7%±2.4%)均低于对照组(55.3%±2.2%),差异有统计学意义(P0.05);3.PCR及Western Blot显示,10μmol/L 5-Aza-Cd R使MDA-MB-231细胞中E-cadherin、WIF1基因表达上调;且呈时间依赖性逐渐升高,而β-catenin的表达随作用时间延长逐渐下降。结论:5-Aza-Cd R能抑制MDA-MB-231细胞增殖,降低其迁移能力,其机制与上调WIF1和E-cadherin的表达并降低β-catenin表达有关。
[Abstract]:Objective: to study the effect of 5-aza-2-deoxycytidine on proliferation and migration of breast cancer MDA-MB-231 cell line. Methods: cell culture and drug treatment: human breast cancer cell line MDA-MB-231 was cultured in vitro. 10 渭 mol/L with different concentrations of 1 渭 mol 路L ~ (-1) O _ (5 渭 mol 路L ~ (-1)) were used. The RPMI1640 medium of 20 渭 mol / L 5-azacytidine 5-Aza-CD R was cultured, and the control group without the drug was set up. 2. The OD value of MDA-MB-231 cell line at different concentrations of 5-Aza-CD R was measured by thiazolyl blue MTT assay, and the change of cell proliferation rate was detected. 3.The migration distance of breast cancer cell line MDA-MB-231 was detected by scratch test. 4. Reverse transcription-polymerase chain reaction was used. E-cadherin was detected by Western Blot after 10 渭 mol/L 5-Aza-CD R treatment for different time. The effects of WIF1 and 尾 -catenin m RNA and protein; 5. SPSS18.0 was used to analyze the difference of cell inhibition rate, expression level of m RNA and protein between treatment group and control group. 5-Aza-CD R inhibited the growth of MDA-MB-231 cells in a dose-and concentration-dependent manner. 2. Cell migration showed that the healing rate of MDA-MB-231 cells in 5 渭 mol / L 10 渭 mol/L drug treated group was 30.0% 卤2.0% after 24 hours of scratch. Compared with the control group (45.0% 卤1.3%), 24. 6% 卤2. 4%) and 5 渭 mol/L after 48 hours. The healing rate of MDA-MB-231 cells in 10 渭 mol/L drug treated group was 37. 3% 卤2. 2% respectively. It was significantly lower than that of the control group (55.3% 卤2.2%, P 0.05). 3. E-cadherin in MDA-MB-231 cells was induced by 10 渭 mol/L 5-Aza-CD R and Western Blot. The expression of WIF1 gene was up-regulated. The expression of 尾 -catenin decreased gradually with the prolongation of time. Conclusion the proliferation of MDA-MB-231 cells can be inhibited by 5-Aza-CD R. The mechanism is related to the up-regulation of WIF1 and E-cadherin expression and the decrease of 尾 -catenin expression.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.9

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