| 贾炳阳,田凯华.5-氮杂-2脱氧胞苷去除p14启动子甲基化对人肺癌细胞生物学功能的影响[J].济宁医学院学报,2019,42(1):1-4 |
| 5-氮杂-2脱氧胞苷去除p14启动子甲基化对人肺癌细胞生物学功能的影响 |
| Effect of 5-Aza-2-deoxycytidine demethylating p14 promoter on biological function in human lung cancer cells |
| 投稿时间：2019-01-20 |
| DOI：10.3969/j.issn.1000-9760.2019.01.001 |
| 中文关键词: p14ARF;DNA甲基化;肺癌 |
| 英文关键词: p14ARF;Lung cancer;DNA Methylation |
| 基金项目:山东省卫生厅科研项目（2007H2035） |
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| 中文摘要: |
| 目的 探讨5-氮杂-2脱氧胞苷（5-aza-2'-deoxycytidine，DAC）去除p14ARF基因启动子异常甲基化状态对人肺癌细胞生物学功能的影响。方法 利用巢式甲基化特异性PCR（NMSP）检测spca1细胞系及beas2b细胞系p14ARF启动子区域甲基化情况，根据细胞类型及是否进行DAC处理将细胞分为4组：spca1 DAC处理组、spca1对照组、beas2b DAC处理组、beas2b对照组，每组设置6个复孔，Western blot检测各组p14ARF蛋白表达情况，流式细胞术进行细胞凋亡检测。结果 NMSP检测到spca1细胞p14启动子区域存在异常的甲基化状态，使用DAC去除p14启动子区异常甲基化spca1细胞中ARF蛋白的表达恢复，细胞凋亡率spca1 DAC处理组（20.69±1.03）%显著高于spca1对照组（10.63±0.82）%（t=13.17，P<0.05）；而在beas2b细胞系以上结果均无明显差异，其中细胞凋亡率beas2b DAC处理组（7.38±0.36）%与beas2b对照组（6.98±0.42）%无显著差异（t=1.25，P>0.05）。结论 DAC去除p14ARF启动子区域异常甲基化后ARF蛋白表达恢复，诱导spca1细胞凋亡，对肺癌发生起到抑制作用。 |
| 英文摘要: |
| Objective To investigate the effect of 5-aza-2'-deoxycytidine (DAC) demethylating the p14ARF gene promoter on the biological function in human lung cancer cells.Methods Nested methylation-specific PCR (NMSP) was used to detect the methylation status of p14ARF promoter region in spca1 cell line and beas2b cell line.According to cell type and DAC treatment,cells were divided into four groups:spca1 DAC treatment group,spca1 control group,beas2b DAC treatment group,beas2b control group,and there were three duplicate wells in each group.The expression of p14ARF protein in each group was detected by Western blot.Apoptosis detection was performed by flow cytometry.Results NMSP detected abnormal methylation status in the p14 promoter region of spca1 cells.The use of DAC to remove aberrant methylation of the p14 promoter region detected the recovery in ARF protein expression in spca1 cells,and the rate of apoptosis was significantly increased.There were significant differences in the experimental group (20.69±1.03)% and control group (10.63±0.82)% (t=13.17,P<0.05).While there was no significant difference in the results of the beas2b cell line,there were no significant differences in the experimental group (7.38±0.36)% and control group (6.98±0.42)% (t=1.25,P>0.05).Conclusion After DAC demethylates the p14ARF promoter region,the expression of ARF protein is recovering,which induces the apoptosis of spca1 cells.So p14 promoter aberrant methylation plays an important role in the development of lung cancer. |
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