干扰高迁移率族蛋白A2的表达对NB4细胞裸鼠种植瘤生长的影响

摘要

目的:建立裸鼠体内 NB4/ShHMGA2 细胞种植瘤模型, 探索稳定敲低高迁移率族蛋白 A2 (HMGA2)的表达对白血病细胞生物学形状的影响. 方法:每组5只4~5周龄的BALB/c裸鼠,经4 Gy 60 Co照射后腋窝皮下分别接种 8 × 106个NB4/ShHMGA2、NB4/ShControl 细胞(对照组),观察成瘤性,3 周后处死裸鼠.免疫组化法检测NB4细胞种植瘤组织切片中Ki-67蛋白的表达水平.结果:NB4/ShHMGA2及对照细胞在裸鼠腋下均成瘤,NB4/ShHMGA2细胞肿瘤生长速度明显慢于对照组,NB4/ShHMGA2 细胞与对照组瘤体体积分别为(1 484.25 ± 156.342)和(3 228.674 ± 285.64)mm3,差异有统计学意义(P < 0.05);NB4/ShHMGA2细胞与对照组瘤块重量分别为(650.46 ± 85.12)和(2 135.33 ± 198.05)mg,差异有统计学意义(P <0.05). 免疫组化检测显示NB4/ShHMGA2细胞组种植瘤组织切片Ki-67蛋白表达水平低于对照组. 结论:敲低HMGA2的表达在体内可抑制白血病细胞的增殖.%Objective To establish a nude mice model for NB4/ShHMGA2 xenograft and explore the effect of HMGA2 knockdown on hematological malignancies. Methods NB4/ShHMGA2 or NB4/ShControl cell lines were established by transfecting the recombinant Lentivirus-HMGA2shRNA and the vacant Lentivirus-NC-marked into NB4 cells. The knockdown of HMGA2 was identified by RT-PCR and Western blot. Ten male BALB/c nude mice aged 4 ~ 5 weeks were equally divided into two groups. The mice irradiated by 4 Gy 60 Co were subcutaneously injected with 8 × 106 NB4/ShHMGA2 or NB4/ShControl cells into one side of axilla. The volumes of xenograft tumor were evaluated using the equation volume (mm3) = (L × W2)/2. The xenograft tumor section was detected by IHC with Ki-67 antibody. Results NB4 cell xenograft tumors developed in all mice of both the two groups. The NB4/ShHMGA2 cells in the nude mice grew at a lower rate than those in the controls. There were statistically significant differences in the volume and weight of xenograft tumor between the two groups [(1 484.25 ± 156.342)mm3 vs (3 228.674 ± 285.64)mm3, P < 0.05] and [(2 135.33 ± 198.05) mg vs (650.46 ± 85.12)mg, P < 0.05]. The Ki-67 protein level in NB4/ShHMGA2 cells xenografts was lower than that in the controls. Conclusion The knockdown of HMGA2 could inhibit proliferation of NB4 cells in NB4 cells xenograft tumor.