BRCA2对乳腺癌干细胞的影响及其机制

BRCA2 affect cancer stem cells in breast cancer and its mechanism analysis

目的分析乳腺癌易感基因2(BRCA2)对乳腺癌干细胞的影响,并且从NOTCH信号通路角度分析BRCA2对乳腺癌干细胞的机制。方法采用已经建立好的BRCA2反转录病毒干扰载体转染成MDA-MB-435S细胞,与此同时在采取球形成试验,检测细胞干扰乳腺癌易感基因2的相关效果,克隆形成试验检测BRCA2下调之后细胞的增殖能力以及蛋白质印迹法检测乳腺癌干细胞中的NOTCH1/NOTCH4在BRCA2中的表达。结果试验组球形成试验的细胞数量为(25.0±1.0)个,对照组球形成试验的细胞数量为(11.0±2.0)个;试验组克隆形成试验的细胞数量为(102.1±12.1)个,对照组克隆形成试验的细胞数量为(50.0±11.1)个;试验组蛋白质印迹法的细胞数量为(75.0±13.1)个,对照组蛋白质印迹法的细胞数量为(42.2±10.0)个,两组数据比较差异有统计学意义(P<0.05);球形成试验以及克隆形成试验的相关结果显示,比较对照组的BRCA2乳腺癌干细胞MDA-MB-435S细胞形成数量明显增加,差异有统计学意义(P<0.05)。蛋白印迹法检测显示,与对照组比较,在沉默的BRCA2乳腺癌干细胞MDA-MB-435S细胞中NOTCH1的表达显著增加,差异有统计学意义(P<0.05),对NOTCH4的表达均未见。结论 BRCA2对乳腺癌干细胞转染成MDA-MB-435S细胞的能力显著增加。

Objective To study the impact of breast cancer susceptibility gene 2（BRCA2） on the breast cancer stem cells, and its mechanism through the analysis of NOTCH signaling pathway. Methods MDA-MB-435 S cells were constructed by the established BRCA2 retroviral vector. The effect BRCA2 gene on the spherical formation ability and stem cell feature of cancer stem cells was determined. The effect of BRCA2 gene down regulation on the cell proliferation ability was measured by colony formation. The effect of NOTCH1 / NOTCH4 on the breast cancer stem cells was analyzed by western blotting. SPSS14.0 statistical software was used for statistical analysis. Results The number of cells in the experimental group ball formation test was（25.0 ±1.0） months, the control group sphere forming cells test was（11.0 ±2.0） months; test group clonogenic cell count test was（102.1 ±12.1） months in the control group the number of colony formation test was（50.0±11.1） months; the number of cells in the experimental group Western blotting was（75.0±13.1）, the number of cells in the control group by Western blotting was（42.2 ±10.0） months, two sets of data comparison the difference was statistically significant（P〈0.05）; sphere forming assay and colony formation associated test results show a comparative control group BRCA2 breast cancer stem cells MDA-MB-435 S cells formed significantly increase the number of data difference was statistically significant（P〈 0.05）; Western blot assay showed that, compared with the control group, in the silence of BRCA2 MDA-MB-435 S cells expressing NOTCH1 significantly increased, the difference was statistically significant（P〈 0.05）, the expression of NOTCH4 average no. Conclusion BRCA2 in breast cancer stem cells transfection into MDA-MB-435 S ability increased significantly.