Xue Jun, Lin Mao Fang
Department of Hematology, The First Affiliated People's Hospotal, Nanjing Medical University, Nanjing 210000, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005 Dec;13(6):969-74.
To explore the regulation mechanism of survivin gene, the NB4 and HL-60 cells were used in experiments, the cell culture in vitro and cell morphological observation were performed and survivin mRNA expression was detected by semi-quantitative RT-PCR. The results showed that the survivin expression in NB4 cell was positive. By treatment of 1 micromol/L ATRA, cell differentiation antigen CD11b was gradually increased ([chi = 47.002, P = 0.000) and CD33 was gradually decreased (chi = 1.614, P = 0.806) with time. Simultaneously, survivin mRNA expression was down-regulated and the cell cycle was arrested at G(0)-G(1) phase (chi = 58.566, P = 0.000). ATRA could down-regulate the survivin mRNA expression of HL-60 cell, but G-CSF, GM-CSF and PHA could up-regulate the survivin expression of HL-60 cell. The cytokine could regulate survivin expression in gene transcription level. The up-regulation of survivin expression was observed while HL-60 cell was stimulated by PHA. The survivin gene expression could be blocked by the survivin antisense oligonucleotide. The survivin mRNA expression of NB4 cell was inhibited by 100 nmol/L-1000 nmol/L survivin antisense oligonucleotide in a dose-dependent manner. The survivin mRNA expression in the NB4 cell was obviously inhibited in 600 nmol/L survivin AS-ODN groups (38%) while the AS-ODN dose increases, the inhibition rate does not descend, but was not inhibited in the control groups, liposomes groups and ODN groups. After NB4 cell was treated by survivin AS-ODN, the typical morphological changes for the apoptosis emerged in NB4 cell. These changes were not found in control groups. It is concluded that PHA, GM-CSF and G-CSF can up-regulate the survivin gene expression, but survivin AS-ODN and ATRA can down-regulate survivin gene expression. The cell cycle arrest at G(0)-G(1) phase while the survivin gene expression was down-regulated by ATRA. It suggested that the survivin gene expression is very related to cell cycle. The morphological changes of cell apoptosis can be observed when the survivin gene expression of NB4 cell was suppressed.
为探讨生存素基因的调控机制，实验采用NB4和HL-60细胞，进行体外细胞培养和细胞形态学观察，并用半定量RT-PCR法检测生存素mRNA表达。结果显示，NB4细胞生存素表达呈阳性。用1μmol/L全反式维甲酸（ATRA）处理后，细胞分化抗原CD11b随时间逐渐升高（χ=47.002，P=0.000），而CD33逐渐降低（χ=1.614，P=0.806）。同时，生存素mRNA表达下调，细胞周期阻滞于G(0)-G(1)期（χ=58.566，P=0.000）。ATRA可下调HL-60细胞生存素mRNA表达，但粒细胞集落刺激因子（G-CSF）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和植物血凝素（PHA）可上调HL-60细胞生存素表达。细胞因子可在基因转录水平调控生存素表达。PHA刺激HL-60细胞时可观察到生存素表达上调。生存素反义寡核苷酸可阻断生存素基因表达。100nmol/L - 1000nmol/L生存素反义寡核苷酸对NB4细胞生存素mRNA表达的抑制呈剂量依赖性。600nmol/L生存素反义寡核苷酸组（38%）对NB4细胞生存素mRNA表达的抑制作用明显，随着反义寡核苷酸剂量增加，抑制率未下降，但对照组、脂质体组和寡核苷酸组均未出现抑制。NB4细胞经生存素反义寡核苷酸处理后，出现典型的凋亡形态学改变，对照组未发现这些改变。结论：PHA、GM-CSF和G-CSF可上调生存素基因表达，但生存素反义寡核苷酸和ATRA可下调生存素基因表达。ATRA下调生存素基因表达时，细胞周期阻滞于G(0)-G(1)期。提示生存素基因表达与细胞周期密切相关。抑制NB4细胞生存素基因表达时，可观察到细胞凋亡的形态学改变。
