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目的观察反义 Bmi-1 对 B 淋巴细胞白血病 Raji 细胞的抑制作用并探讨其机制。方法在体外转染中通过阳离子脂质体法将反义质粒导入 Raji 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 Raji 细胞增生的抑制作用;利用流式细胞仪分析细胞周期;采用免疫荧光组化法检测反义 Bmi-1对 Raji 细胞的 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,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,空白对照组细胞集落数为(89.7±8.07)/10~3细胞,空质粒组细胞集落数为(81.3±6.91)/10~3细胞,转染反义质粒组细胞集落数为(50.0±5.21)/10~3细胞(P<0.01);与对照组细胞相比反义组 G_1期所占比例明显上升(P<0.05);p16蛋白表达也明显增强。结论反义 Bmi-1对 Raji 细胞的体外生长具有明显的抑制作用。
Objective To investigate the inhibitory effect of antisense Bmi-1 on Raji cells of B lymphoblastic leukemia and its mechanism. Methods Antisense plasmids were transfected into Raji cells by cationic liposome method in vitro and positive clones were screened by G418. The antisense Bmi-1 expression plasmid was used to detect the proliferation of Raji cells by MTT assay and colony formation assay in vitro The effect of antisense Bmi-1 on the up-regulation of p16 protein in Raji cells was detected by immunofluorescence staining. Results Compared with control group (blank control group and empty plasmid group), the expression of antisense Bmi-1 expression plasmid group was significantly slower than that of control group (P <0.05). The expression of antisense Bmi-1 plasmid group and control group (blank control and empty plasmid (89.7 ± 8.07) / 10 ~ 3 cells in the blank control group and (81.3 ± 6.91) / 10 ~ (-3) cells in the empty plasmid group compared with the control group 3 cells. The number of colony of transfected antisense plasmid group was (50.0 ± 5.21) / 10 ~ 3 cells (P <0.01), and the proportion of G_1 phase in antisense group was significantly higher than that of control cells (P <0.05) ; p16 protein expression was also significantly enhanced. Conclusion Antisense Bmi-1 can significantly inhibit the growth of Raji cells in vitro.