Objective To investigate the effects of c⁃myc promoter binding protein 1(MBP⁃1)gene on the proliferation of human Saos⁃2 osteo⁃sarcoma cells in vitro. Methods Saos⁃2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP⁃1 shRNA). Two MBP⁃1 shRNA sequences and one neg⁃ative control shRNA sequence were designed ,synthesized and cloned into pSIREN⁃retroQ plasma. Then the recombinant plasmids were construct⁃ed and transfected into human Saos⁃2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP⁃1 mRNA and protein in Saos⁃2 cells were detected by real⁃time PCR and Western blot ,respectively. The effects of altered expression of MBP⁃1 on cell proliferation were measured by CCK⁃8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos⁃2 were determined by Western blot. Results PCR and sequenc⁃ing results indicated that the recombinant plasmids pSIREN⁃retroQ was constructed. The relative expression level of MBP⁃1 mRNA in the MBP⁃1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex⁃pression levels of MBP⁃1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos⁃2 cells at 48,72,and 96 hours after MBP⁃1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con⁃trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP⁃1 gene promotes the proliferation of human Saos⁃2 osteosarcoma cells. MBP⁃1 gene may become the new tar⁃get of gene therapy for osteosarcoma.%目的：探讨c⁃myc启动子结合蛋白1（MBP⁃1）基因沉默对人骨肉瘤Saos⁃2细胞生长的影响。方法实验分为3组：正常对照组（未转染骨肉瘤细胞）、阴性对照组（转染错义序列组）和沉默组（转染MBP⁃1 shRNA组）。设计2条MBP⁃1基因的RNA干扰片段及1条阴性对照siRNA，并与pSIREN⁃retroQ质粒连接。将重组pSIREN⁃retroQ质粒通过Lipofectamine 2000脂质体转染骨肉瘤Saos⁃2细胞。实时PCR和Western blot分别检测MBP⁃1表达。CCK⁃8法对MBP⁃1沉默后骨肉瘤Saos⁃2细胞生长进行检测。Western blot检测对照组和沉默组cyclin D1和cyclin E的表达。结果通过PCR扩增及测序，说明已成功构建MBP⁃1沉默及对照重组pSIREN⁃retroQ 质粒。实时PCR结果显示，沉默组MBP⁃1 mRNA相对表达量与对照组相比显著下调（P<0.05）。Western blot结果显示，沉默组MBP⁃1蛋白表达量与对照组相比也显著下调。CCK⁃8法结果表明，沉默组细胞在48、72和96 h时增殖能力均比对照组显著升高（P<0.05）。沉默组cyclin D1和cyclin E的表达显著高于对照组（P<0.05）。结论 MBP⁃1基因沉默后骨肉瘤Saos⁃2细胞生长被明显促进，为寻找骨肉瘤基因治疗新靶点打下基础。
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