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目的:采用短发夹状RNA(short hairpin RNA,shRNA)干扰技术沉默乳腺癌细胞MDA-MB-231中转移相关基因(metastasis-associated gene 1,MTA1)的表达,并观察其对15-脂氧合酶-2(15-lipoxygenase-2,15-LOX-2)、p53及bcl-2表达的影响。方法:将shRNA-MTA1载体质粒稳定转染MDA-MB-231细胞,MTT法检测细胞增殖抑制情况,FCM法检测细胞周期及凋亡情况,RT-PCR和Western印迹法检测MTAl、15-LOX-2、p53及bcl-2 mRNA和蛋白表达情况。结果:shRNA-MTA1能明显降低MTA1基因在MDA-MB-231细胞中的表达量;抑制MDA-MB-231细胞的增殖,诱导其凋亡,并使细胞被阻滞在G1期,与对照组相比,差异有统计学意义(P<0.01)。转染shRNA-MTA1可使MDA-MB-231细胞中15-LOX-2和p53 mRNA及蛋白的表达明显增强(P<0.01),而MTA1和bcl-2 mRNA表达明显减弱(P<0.01)。结论:MTA1基因可抑制乳腺癌MDA-MB-231细胞的增殖,诱导其凋亡,该作用可能与上调细胞中15-LOX-2和p53的表达,下调bcl-2的表达有关。
OBJECTIVE: To silence the expression of metastasis-associated gene 1 (MTA1) in breast cancer cell line MDA-MB-231 by short hairpin RNA (shRNA) interference technique and observe its effect on 15-lipoxygenase 5-lipoxygenase-2, 15-LOX-2, p53 and bcl-2. Methods: The shRNA-MTA1 plasmid was stably transfected into MDA-MB-231 cells. The cell proliferation was determined by MTT assay. Cell cycle and apoptosis were detected by FCM. The expressions of MTA1 and 15-LOX- 2, p53 and bcl-2 mRNA and protein expression. Results: shRNA-MTA1 could significantly decrease the expression of MTA1 in MDA-MB-231 cells, inhibit the proliferation of MDA-MB-231 cells and induce the apoptosis of cells, Compared with the difference was statistically significant (P <0.01). The expression of 15-LOX-2 and p53 mRNA and protein in MDA-MB-231 cells transfected with shRNA-MTA1 was significantly increased (P <0.01), while the expression of MTA1 and bcl-2 mRNA was significantly decreased (P <0.01). CONCLUSION: MTA1 can inhibit the proliferation and induce the apoptosis of breast cancer MDA-MB-231 cells, which may be related to the up-regulation of 15-LOX-2 and p53 expression and the down-regulation of bcl-2 expression.