METHODS: DUSP6-siRNA was used to inhibit the expression of DUSP6 directly and miR-145 to inhibit the expression of DUSP6 either in MDA-MB-231 breast cancer cells and successful transfection being confirmed by Real-time PCR and Western Blotting. Down regulation of DUSP6 in MDA-MB-231 cells suppressed the cell proliferation as investigated by MTT assay and colony form assay. Transwell test and Scratch assay were conducted to investigate the migration and invasion of MDA-MB-231 cells. T-test (two-tailed) was used to compare differences between groups, and the significance level was set at P<0.05.
RESULTS: DUSP6 mRNA expression and protein expression were reduced after transfection with DUSP6-siRNA directly and similar trend with transfection with miR-145. The treated group with DUSP6-siRNA or miR-145 suppressed MDA-MB-231 cells proliferation, migration and invasion, and meanwhile the cells were arrested at G0/G1 phase.
CONCLUSIONS: DUSP6 plays a role in triple-negative breast cancer cells that might promote growth in MDA-MB-231 triple-negative breast cancer cells.
方法:用DUSP6-siRNA直接抑制MDA-MB-231乳腺癌细胞中DUSP6的表达,用miR-145抑制DUSP6的表达,并通过Real-timePCR和WesternBlotting证实转染成功。如通过MTT测定和集落形式测定所研究的,MDA-MB-231细胞中DUSP6的下调抑制了细胞增殖。进行Transwell试验和划痕试验以研究MDA-MB-231细胞的迁移和侵袭。T检验(双尾)用于比较组间差异,显著性水平为P<0.05。
结果:直接转染DUSP6-siRNA后,DUSP6mRNA表达和蛋白表达降低,与转染miR-145的趋势相似。DUSP6-siRNA或miR-145治疗组抑制MDA-MB-231细胞增殖,移民和入侵,同时细胞被阻滞在G0/G1期。
结论:DUSP6在三阴性乳腺癌细胞中发挥作用,可能促进MDA-MB-231三阴性乳腺癌细胞的生长。