ECT2基因沉默调控人乳腺癌细胞增殖和凋亡及其机制探讨

发布时间：2018-05-20 18:28

本文选题：乳腺肿瘤 + 基因沉默　；参考：《肿瘤》2017年06期

【摘要】：目的:探讨上皮细胞转化序列2癌基因(epithelial cell transforming sequence 2 oncogene,ECT 2)沉默对乳腺癌细胞增殖和凋亡的影响,以及其潜在的作用机制。方法:首先采用实时荧光定量PCR和蛋白质印迹法检测正常乳腺上皮细胞(MCF-10A)和乳腺癌细胞(MDA-MB-231、SK-BR-3、MCF-7和BT474)中ECT2 mRNA及蛋白的表达水平。将特异性靶向沉默ECT 2基因的siRNA(ECT2 siRNA)转染至乳腺癌MDA-MB-231和MCF-7细胞,并用细胞外调节蛋白激酶(extracellular regulated protein kinase,ERK)通路激活剂处理或者微RNA-101(microRNA-101,miR-101)抑制子转染后,采用CCK-8和FCM法分别检测细胞增殖和凋亡情况,采用实时荧光定量PCR和蛋白质印迹法检测细胞中磷酸化ERK(phospho-ERK,p-ERK)、Ras相关的C3肉毒底物1(Ras-related C3 botulinum toxin substrate 1,Rac1)和miR-101的表达水平。结果:与正常乳腺上皮细胞相比,乳腺癌细胞中ECT2 mRNA和蛋白的表达水平均明显升高(P值均0.05)。ECT 2基因沉默后,乳腺癌细胞增殖被明显抑制(P0.05),细胞凋亡被明显促进(P0.05),并且细胞中p-ERK和Rac1表达被明显下调(P值均0.05),而miR-101表达被明显上调(P0.05)。ERK激活剂作用后,ECT2 siRNA转染对p-ERK、miR-101和Rac1表达的影响被明显反转(P值均0.05);miR-101抑制子转染后,ECT2 siRNA转染对细胞增殖、凋亡以及Rac1表达的影响被反转(P值均0.05),而对p-ERK表达没有明显影响(P0.05)。结论:ECT 2基因沉默可能通过ERK/miR-101/Rac1信号转导通路,调节乳腺癌细胞的增殖和凋亡。
[Abstract]:Aim: to investigate the effect of epithelial cell transforming sequence 2 oncogeneECT 2 silencing on the proliferation and apoptosis of breast cancer cells and its potential mechanism. Methods: the expression levels of ECT2 mRNA and protein in normal breast epithelial cells (MCF-10A) and breast cancer cells (MDA-MB-231SK-BR-3MF-7 and BT474) were detected by real-time fluorescence quantitative PCR and Western blot. The specific siRNA(ECT2 siRNAs targeting silenced ECT 2 gene were transfected into breast cancer MDA-MB-231 and MCF-7 cells, then were treated with extracellular regulated protein kinase activator or microRNA-101 miR-101) suppressor transfection. CCK-8 and FCM were used to detect cell proliferation and apoptosis, and real-time fluorescence quantitative PCR and Western blot were used to detect the expression of 1(Ras-related C3 botulinum toxin substrate 1 (Rac1) and miR-101. Results: compared with normal breast epithelial cells, the expression of ECT2 mRNA and protein in breast cancer cells increased significantly after the silencing of 0.05).ECT 2 gene. The proliferation of breast cancer cells was significantly inhibited, the apoptosis was significantly promoted, and the expression of p-ERK and Rac1 were significantly down-regulated (P = 0.05), while the expression of miR-101 was up-regulated by P0.05. ERK activator, and the expression of p-ERKT2-miR-101 and Rac1 was significantly up-regulated by siRNA transfection with ECT2. The proliferation of cells was induced by siRNA transfection of ECT2 siRNA after transfection with 0.05mmiR-101 suppressor. The effect of apoptosis and Rac1 expression was reversed P value was 0.05, but no significant effect on p-ERK expression P0.05. Conclusion the cell proliferation and apoptosis of breast cancer cells may be regulated by ERK/miR-101/Rac1 signal transduction pathway.
【作者单位】：江西省萍乡市人民医院乳腺外科;
【分类号】：R737.9

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