【摘要】 目的构建含表皮生长因子受体2基因(HER2)的短发夹状双链RNA(shRNA)重组质粒,体外观察对人乳腺癌细胞SKBr3的HER2 mRNA及蛋白表达的影响。方法利用分子克隆技术,将含HER2的双链DNA,与经双酶切后的载体pSilencer连接,构建pSilencer-HER2重组质粒,在脂质体的介导下转染高表达HER2的乳腺癌细胞系SKBr3。RT-PCR分析HER2 mRNA的表达,Western blot检测蛋白的表达,MTT法测定细胞增殖情况。结果酶切鉴定证实重组质粒构建成功,转染后能明显抑制HER2 mRNA及蛋白的表达,抑制细胞的增殖。结论构建的pSilencer-HER2重组质粒能有效地降低人乳腺癌细胞HER2的表达,抑制细胞的增殖,为HER2高表达预后不良的乳腺癌基因治疗提供新策略。更多还原

【Abstract】 Objective To construct a expression vector of short hairpin RNA of human epidermal growth factor receptor gene(HER2) and observe its effects on HER2 expression and cell growth of human breast cancer cell line SKBr3 in vitro.Methods The recombinant plasmid pSilencer-HER2 was constructed by combination of double-strand DNA containing HER2 with vector pSilencer that underwent restriction digest.Then these recombinant plasmids were transfected by liposome into SKBr3 cells that highly express HER2.RT-PCR was used to analyze HER2 mRNA expression,Western-blot to detect its protein product and MTT to assess cell proliferation.Results Restriction enzyme digesting assay confirms successful construction of recombinant plasmid pSilencer-HER2.And its transfection into human breast cancer cells can significantly reduce expression of HER2 mRNA and its protein product,thus inhibit cell proliferation.Conclusion Recombinant plasmid pSilencer-HER2 can significantly reduce HER2 expression of human breast cancer cell SKBr3 and inhibit its growth.The results obtained provide a new strategy for gene therapy for breast cancers with high HER2 expression and improved poor prognosis.更多还原