目的观察反义Bmi-1表达质粒在体外对T淋巴细胞白血病Jurkat细胞的生长抑制作用。方法选择Bmi-1基因起始密码子及与锌指结构对应的171bp核苷酸片段,反向连接于pLNCX2质粒上,得到反义Bmi-1表达质粒;通过脂质体转染将质粒导入Jurkat细胞中,以G418进行筛选得到阳性克隆;以MTT法和体外集落形成实验检测反义Bmi-1表达质粒对Jurkat细胞增殖的抑制作用;用流式细胞术分析细胞周期;采用免疫荧光组化法检测反义Bmi-1对Jurkat细胞P16蛋白的上调作用。结果转染反义Bmi-1表达质粒组与对照组(空白对照及空质粒组)相比较,生长速度明显变慢,克隆形成能力明显下降,空白对照组Jurkat细胞集落数为(90.70±9.07)/103细胞,空质粒组Jurkat细胞集落数为(83.30±6.11)/103细胞,转染反义质粒组Jurkat细胞集落数为(56.00±5.56)/103细胞(P<0.01);P16蛋白表达也明显增强。结论反义Bmi-1对Jurkat细胞的体外生长具有明显的抑制作用,并上调了P16蛋白的表达。 Objective To observe the inhibitory effect of antisense Bmi-1 expression plasmid on T lymphocyte leukemia Jurkat cell growth in vitro. METHODS: The Bmi-1 gene start codon and the 171 bp nucleotide fragment corresponding to the zinc finger structure were selected and reversely ligated to the pLNCX2 plasmid to obtain the antisense Bmi-1 expression plasmid; the plasmid was introduced into Jurkat by liposome transfection. In the cells, positive clones were screened with G418; the inhibition of antisense Bmi-1 expression plasmid on the proliferation of Jurkat cells was detected by MTT assay and in vitro colony formation assay; cell cycle was analyzed by flow cytometry; immunofluorescence staining method was used. The upregulation of P16 protein in Jurkat cells by antisense Bmi-1 was detected. Results Compared with the control group (blank control and empty plasmid group), the antisense Bmi-1 expression plasmid group transfected with the antisense Bmi-1 expression plasmid showed a significantly slower growth rate and decreased colony formation ability. The number of colony-forming Jurkat cells in the blank control group was (90.70±9.07). The colony number of Jurkat cells in (103) cells and empty plasmid group was (83.30±6.11)/103 cells, and the colony number of Jurkat cells transfected with antisense plasmid group was (56.00±5.56)/103 cells (P<0.01); the expression of P16 protein was also Obviously enhanced. Conclusion Antisense Bmi-1 has obvious inhibitory effect on the growth of Jurkat cells in vitro and up-regulates the expression of P16 protein.