沉默CXCL1基因抑制三阴性乳腺癌细胞MDA-MB-231的迁移、侵袭的研究

为探讨CXCL1基因对三阴性乳腺癌细胞MDA-MB-231的迁移、侵袭作用的影响,该研究设计针对CXCL1基因的小干扰RNA,用实时荧光定量PCR和酶联免疫吸附测定试验分别在RNA和蛋白质水平上检测干扰效率;采用流式细胞术学技术检测细胞的周期和凋亡情况;采用transwell迁移和侵袭试验分别检测细胞的迁移及侵袭能力。结果显示,与siRNA-NC组细胞相比,siCXCL1-1、siCXCL1-2、siCXCL1-3细胞中CXCL1基因的mRNA和蛋白表达均下调,且si CXCL1-3干扰效率最高,mRNA及蛋白表达水平分别降低了75%(p<0.01)和46%(p<0.01)。细胞凋亡试验和细胞周期试验结果显示,沉默CXCL1基因后对三阴性乳腺癌细胞系MDA-MB-231细胞的凋亡和周期无明显影响,差异无统计学意义(p>0.05)。transwell小室迁移和侵袭试验显示,沉默CXCL1基因能够显著抑制三阴性乳腺癌细胞MDA-MB-231的迁移和侵袭能力(p<0.01)。研究成果为临床乳腺癌中以CXCL1基因为靶点的分子治疗提供了理论依据。

Study on Inhibitory Effect of Silence Gene CXCL1 on Migration/Invasion of Triple Negative Breast Cancer Cell MDA-MB-231

To investigate the effect of CXCL1 gene on migration/invasion of triple negative breast cancer cell MDA-MB-231, small interference RNA(siRNA) targeting the CXCL1 gene was designed and the interference efficiencies were detected at the RNA and protein levels using real-time fluorescent quantitative PCR and ELISA in this study. The cell cycle and apoptosis were analyzed by flow cytometry. Cellular migration and invasion abilities were detected by transwell migration and invasion assay. The results showed that compared with siRNA-NC group,the expression of CXCL1 m RNA and protein levels were down-regulated in si CXCL1-1, si CXCL1-2 and si CXCL1-3cells, and the most efficient interference RNA was si CXCL1-3, with the expression of m RNA and protein decreased by 75%(p<0.01) and 46%(p<0.01). The results of cell apoptosis assay and cell cycle assay indicated that silencing CXCL1 gene had no significant effect on the cell apoptosis and cycle of triple negative breast cancer cell line MDA-MB 231 and there was no statistical significance(p>0.05). Transwell chamber migration and invasion assay showed that silence CXCL1 gene could significantly inhibit the migration and invasion abilities of triple negative breast cancer cell MDA-MB 231(p<0.01). The research showed that the expression of silence CXCL1 gene could inhibit the migration and invasion abilities of triple negative breast cancer cell MDA-MB 231, but had no significant effect on cell cycle and apoptosis.