下调星形细胞上调基因1表达对乳腺癌细胞凋亡的影响
| 作者: |
1常亮,
1户庄,
1周振宇,
1付传恒,
1马明德
1 河南大学淮河医院 甲状腺乳腺外科，河南 开封 475000 |
| 通讯: | |
| DOI: | 10.3978/.2018.05.008 |
摘要
目的：探讨下调星形细胞上调基因1（AEG-1）对乳腺癌细胞凋亡的影响。
方法：通过脂质体介导的方法，将乳腺癌MCF-7细胞分别转染AEG-1 siRNA（AEG-1 siRNA）和阴性对照siRNA（阴性对照组），以无处理的MCF-7细胞为空白对照组。验证转染效果后，分别采用流式细胞术与Western blot检测各组细胞的凋亡以及凋亡相关蛋白caspase-3和caspase-9的表达。
结果：与空白对照组细胞比较，AEG-1 siRNA组MCF-7细胞AEG-1蛋白表达明显降低、细胞凋亡率明显升高、caspase-3和caspase-9蛋白表达明显上调（均P<0.05）；阴性对照组与空白对照组间以上各项指标均无统计学差异（均P>0.05）。
结论：下调乳腺癌细胞AEG-1的表达能促进caspase-3和caspase-9的表达，从而诱导乳腺癌细胞凋亡，因此，AEG-1在乳腺癌细胞中可能起了抑制细胞凋亡的作用，机制可能与其调节凋亡相关蛋白的表达有关。
关键词：
乳腺肿瘤；星形细胞上调基因1；细胞凋亡；凋亡调节蛋白质类
方法：通过脂质体介导的方法，将乳腺癌MCF-7细胞分别转染AEG-1 siRNA（AEG-1 siRNA）和阴性对照siRNA（阴性对照组），以无处理的MCF-7细胞为空白对照组。验证转染效果后，分别采用流式细胞术与Western blot检测各组细胞的凋亡以及凋亡相关蛋白caspase-3和caspase-9的表达。
结果：与空白对照组细胞比较，AEG-1 siRNA组MCF-7细胞AEG-1蛋白表达明显降低、细胞凋亡率明显升高、caspase-3和caspase-9蛋白表达明显上调（均P<0.05）；阴性对照组与空白对照组间以上各项指标均无统计学差异（均P>0.05）。
结论：下调乳腺癌细胞AEG-1的表达能促进caspase-3和caspase-9的表达，从而诱导乳腺癌细胞凋亡，因此，AEG-1在乳腺癌细胞中可能起了抑制细胞凋亡的作用，机制可能与其调节凋亡相关蛋白的表达有关。
Effect of down-regulating the expression of astrocyte elevated gene 1 on apoptosis of breast carcinoma cells
Abstract
Objective: To investigate effect of down-regulating the expression of astrocyte elevated gene 1 (AEG-1) on cell apoptosis in breast cancer cells.
Methods: Through liposome-mediated method, the breast cancer MCF-7 cells were transfected with AEG-1 siRNA (AEG-1 siRNA group) or negative control siRNA (negative control group), and the untreated MCF-7 cells served as blank control group. The transfection effects were observed, and then the apoptosis and the expressions of the apoptosis-associated protein caspase-3 and caspase-9 in each group of cells were determined by flow cytometry and Western blot analysis, respectively.
Results: In AEG-1 siRNA group compared with blank control group, the expression of AEG-1 protein was significantly decreased, apoptosis rate was significantly increased, and the expressions of caspase-3 and caspase-9 were significantly up-regulated (all P<0.05). No significant differences were noted in above indexes between negative control group and blank control group (all P>0.05).
Conclusion: Down-regulating AEG-1 expression can promote the expressions of caspase-3 and caspase-9, and thereby induce the apoptosis of the breast cancer cells. Thus, AEG-1 may play a role in apoptosis inhibition in breast cancer cells, and the mechanism is probably related to its regulating the expressions of apoptosis-associated proteins.
Keywords：
Breast Neoplasms; Astrocyte Elevated Gene 1; Apoptosis; Apoptosis Regulatory Proteins
Methods: Through liposome-mediated method, the breast cancer MCF-7 cells were transfected with AEG-1 siRNA (AEG-1 siRNA group) or negative control siRNA (negative control group), and the untreated MCF-7 cells served as blank control group. The transfection effects were observed, and then the apoptosis and the expressions of the apoptosis-associated protein caspase-3 and caspase-9 in each group of cells were determined by flow cytometry and Western blot analysis, respectively.
Results: In AEG-1 siRNA group compared with blank control group, the expression of AEG-1 protein was significantly decreased, apoptosis rate was significantly increased, and the expressions of caspase-3 and caspase-9 were significantly up-regulated (all P<0.05). No significant differences were noted in above indexes between negative control group and blank control group (all P>0.05).
Conclusion: Down-regulating AEG-1 expression can promote the expressions of caspase-3 and caspase-9, and thereby induce the apoptosis of the breast cancer cells. Thus, AEG-1 may play a role in apoptosis inhibition in breast cancer cells, and the mechanism is probably related to its regulating the expressions of apoptosis-associated proteins.