过表达外源M-CSF促进MCF7细胞增殖

Overexpression of M-CSF Promotes Proliferation of MCF7 Cells

为探讨过表达外源巨噬细胞集落刺激因子(M-CSF)对细胞增殖的影响,将重组载体pCMV/cyto/myc-M-CSF转染MCF7细胞、G418筛选,RT-PCR、Western印迹和免疫荧光鉴定M-CSF的mRNA表达、蛋白表达及定位,通过计算细胞倍增时间、MTT法及反义寡核苷酸抑制实验分析M-CSF对细胞增殖的影响.结果表明,转染M-CSF的MCF7细胞过表达M-CSF mRNA及蛋白,并且定位表达于细胞质;与转染空载体及未转染M-CSF组细胞比较,转染M-CSF的MCF7细胞倍增时间明显缩短、增殖能力显著增强,M-CSF的特异性反义寡核苷酸能抑制转染M-CSF的MCF7细胞的增殖,且抑制率随着反义寡核苷酸浓度的增高而增强;以上结果提示,胞质过表达M-CSF可促进MCF7细胞的增殖.

To explore the effect of macrophage colony stimulating factor （M-CSF） on the cell proliferation, pCMV/cyto/myc-M-CSF vectors were transfected into MCF7 cells using liposomes and screened with G418. RT-PCR, Western blot and immunofluorescence assays were performed to verify the M-CSF mRNA or protein expressions and localization. The results indicated that a high level of M-CSF expression in the cytoplasm was detected which promoted cell proliferation as shown by cell doubling time, MTT assays in M-CSF-transfected MCF7 cells. In addition, the proliferation of MCF7 cells transfected with M-CSF-specific antisense oligonucleotides was significantly inhibited than those of pCMV/cyto/myc transfected or untransfected MCF7 cells. We conclude that the overexpression of M-CSF promotes the proliferation of MCF7 cells.