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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 analyze the effect of breast cancer susceptibility gene 2 (BRCA2) on breast cancer stem cells and to analyze the mechanism of BRCA2 on breast cancer stem cells from the NOTCH signaling pathway. Methods The established BRCA2 retroviral vector was transfected into MDA-MB-435S cells. At the same time, the effect of cell-mediated interference on breast cancer susceptibility gene 2 was detected by sphere formation assay. The clonogenic assay was used to detect the expression of BRCA2 The expression of NOTCH1 / NOTCH4 in breast cancer stem cells in BRCA2 was detected by Western blotting after proliferation. Results The number of cells in the sphere formation test was (25.0 ± 1.0) and that in the control group was (11.0 ± 2.0). The number of cells in the experiment group was (102.1 ± 12.1) (50.0 ± 11.1). The number of cells in the experimental group was (75.0 ± 13.1) and the number of cells in the control group was (42.2 ± 10.0). The two groups of data (P <0.05). The results of spheroid formation test and colony formation assay showed that the numbers of MDA-MB-435S cells in BRCA2 breast cancer stem cells were significantly increased compared with the control group (P <0.05). Western blotting showed that NOTCH1 expression was significantly increased in the silenced BRCA2 breast cancer stem cells MDA-MB-435S cells compared with the control group, the difference was statistically significant (P <0.05), not the expression of NOTCH4. Conclusion The ability of BRCA2 to transfect breast cancer stem cells into MDA-MB-435S cells is significantly increased.