摘要
目的:探讨RNA干扰(RNAi)技术沉默胰岛素生长因子结合蛋白2(IGFBP-2)对乳腺癌细胞株MCF-7增殖的影响。方法:构建IGFBP-2序列特异性siRNAs重组表达质粒pSilencerTM2.1-U6 neo-IGFBP-2,转染MCF-7细胞,采用RT-PCR检测重组质粒对IGFBP-2 mRNA表达的影响,MTT法观察重组质粒对MCF-7细胞体外生长的移植作用,流式细胞仪检测重组质粒对MCF-7细胞凋亡的作用。结果:成功构建了pSilencerTM2.1-U6 neo-IGFBP-2重组质粒,并成功转染MCF-7细胞;重组质粒抑制IGFBP-2的mRNA表达近70%,转染重组质粒72 h后MCF-7细胞的增殖活力降低为(55.8±3.7)%,细胞凋亡率为42.55%。结论:重组质粒pSilencerTM2.1-U6 neo-IGFBP-2明显减低IGFBP-2在MCF-7细胞中的表达,抑制MCF-7细胞增殖,促进MCF-7细胞凋亡。
Objective： To investigate the effects of RNA interference to IGFBP -2 gene on proliferation of the breast cancer cell line MCF - 7. Methods： A expression vector for pSilencerTM 2. 1 - U6 neo was used to carry out siRNA. The plasmid pSilencerTM 2. 1 - U6 neo - IGFBP - 2 was constructed and confirmed by restriction cutting. The plasmid pSilencerTM 2. 1 - U6 neo - IGFBP - 2 was transferred in- to MCF - 7 cells. After MCF - 7 cells was transfected for 72 h, the expression of IGFBP - 2 was detected by RT - PCR method. MT and FCM were used to observe the proliferation and apoptosis of MCF - 7 cells. Results ： pSilencerTM 2. 1 - U6 neo - IGFBP - 2 was constructed and transfected into MCF - 7 ceils successfully. The expression of IGFBP - 2 mRNA in MCF - 7 cells transfected with plasmid pSilencerTM 2. 1 - U6 neo - IGFBP - 2 decreased nearly 70% ; the plasmid pSileneerTM 2. 1 - U6 neo - IGFBP - 2 could inhibit the growth of MCF - 7 cells to （55.8 ± 3.7） % and induce the apoptosis of MCF - 7 cells about 42. 55 %. Conclusion ： siRNA could significantly inhibit IGFBP - 2 expression , suppress growth of MCF -7 cells and induce apoptosis of MCF -7 cells.
出处
《中国妇幼保健》
CAS
北大核心
2013年第28期4719-4721,共3页
Maternal and Child Health Care of China
基金
吉林省科技厅资助项目〔201015220〕
吉林省科技厅重点实验室项目〔20122113〕
