【摘要】 目的利用小干扰RNA（small interfering RNA,siRNA）下调VEGFR-1基因的表达研究对乳腺癌增殖的影响。方法设计针对人VEGFR-1基因的小干扰RNA,直接化学合成,脂质体Lipofectamine^TM2000作为转染试剂,转染人乳腺癌细胞系MCF-7,然后四甲基偶氮唑蓝（MTT）法检测细胞增殖变化,半定量RT-PCR和蛋白印迹试验检测VEGFR-1 mRNA和蛋白的表达,Hoechst33258荧光染色分析细胞凋亡。结果实验结果显示:下调VEGFR-1基因表达,乳腺癌细胞MCF-7增殖被抑制,VEGFR-1 mRNA和其蛋白的表达明显降低,Hoechst33258荧光染色显示转染siRNA72h后MCF-7细胞凋亡明显增加。结论化学合成的siRNA能成功下调VEGFR-1基因的表达,并抑制乳腺癌细胞MCF-7的增殖、增加凋亡;下调VEGFR-1基因的表达有可能是潜在的肿瘤治疗方法。更多还原

【Abstract】 Objective To investigate the influence of down-regulating VEGFR-1 gene expression on proliferation of breast cancer cells through siRNA.Methods Design human VEGFR-1 siRNA and synthesize chemically.The siRNA was transfected into human MCF-7 cells using cationic liposome LipofectamineTM 2000 to down-regulate VEGFR-1 gene.Proliferation of cells was measured by MTT assay,Changes of VEGFR-1 mRNA and protein expression were analyzed by reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting,the cells were observed with an invert microscope in shape and with a fluorescence microscope in apoptosis dyed by Hoechst33258.Results The results of experiments showed that siRNA effectively down-regulated VEGFR-1 gene,inhibited the proliferation of human MCF-7 and markedly reduced the expression of VEGFR-1 mRNA and protein.Seventy-two hours after siRNA being transfected,apoptosis obviously increased in stained cells by Hoechst33258 under fluorescent microscope.Conclusion The siRNA by chemical synthesis can efficiently down-regulate VEGFR-1 gene expression,inhibit MCF-7 proliferation of breast cancer cells and increse apoptosis.Down-regulating VEGFR-1 gene may have the potential as a therapeutic modality of human cancer.更多还原