Ad-NLS-RARα对HL-60细胞增殖及ATRA诱导的HL-60细胞分化的影响及其机制

Effect of Recombinant Adenovirus Carrying NLS-RARα Gene on the Proliferation of HL-60 Cell and the

中文关键词:  重组腺病毒 NLS-RARα HL-60细胞 ATRA 细胞增殖 细胞分化

英文关键词:Recombined adenovirus NLS-RARα HL-60 cells ATRA Cell proliferation Cell differentiation

中文摘要:

      目的 将已构建验证成功的重组腺病毒Ad-NLS-RARα感染至HL-60细胞中，探讨其对HL-60细胞增殖及全反式维甲酸（ATRA）诱导的HL-60细胞分化的影响及机制。方法 将验证正确的腺病毒经Protamine sulfate辅助感染HL-60细胞，检测感染效率，用RT-PCR和Western blot检测目的基因在HL-60细胞中的表达情况。MTT法检测细胞增殖情况。病毒感染HL-60细胞24 h后，用2.5 μmol/L ATRA处理，流式细胞术（FCM）检测细胞表面分化抗原CD11b的表达。RT-PCR和Western blot检测C-MYC基因在mRNA和蛋白水平的表达。结果 在Protamine sulfate辅助感染作用下，重组腺病毒Ad-NLS-RARα和阴性对照腺病毒Ad-KZ对HL-60细胞的感染效率可达70%～80%。NLS-RARα基因的RT-PCR和Western blot结果显示，感染了重组腺病毒Ad-NLS-RARα的HL-60细胞的NLS-RARα基因的mRNA及蛋白表达水平明显增高（P<0.05）。MTT法检测表明感染了重组腺病毒Ad-NLS-RARα的细胞增殖能力增高 (P<0.05)。FCM检测结果显示,经ATRA处理后，感染Ad-NLS-RARα重组腺病毒的HL-60细胞，CD11b表达较阴性对照组和未感染组下调（P<0.05）。C-MYC基因的RT-PCR和Western blot检测结果表明，经ATRA 处理后，感染Ad-NLS-RARα重组腺病毒的细胞C-MYC基因的mRNA和蛋白水平均高于其他两组(P<0.05)。结论 重组腺病毒Ad-NLS-RARα可以促进HL-60细胞的增殖，并通过上调C-MYC基因的表达，抑制ATRA诱导的HL-60细胞分化。

英文摘要:

      Objective To explore the effect and mechanism of recombined adenovirus carrying NLS-RARα gene on proliferation of HL-60 cells and the differentiation of HL-60 cells induced by ATRA. Methods HL-60 cells was infected with Ad-NLS-RARα and control virus Ad-KZ. The efficiency of infection was detected by FCM. The mRNA and protein levels of NLS-RARα were assessed by Real-time PCR (RT-PCR) and Western blot, respectively. MTT assay were applied to determine proliferation of HL-60 cells. Cell surface differentiation antigen CD11b of infected HL-60 cell induced by ATRA was examined by FCM. The mRNA and protein levels of C-MYC of infected HL-60 cell induced by ATRA were determined by Real-time PCR (RT-PCR) and Western blot assay. Results The efficiency of infection of Ad-NLS-RARα and Ad-KZ on HL-60 cell was 70%-80%. The mRNA and protein levels of NLS-RARα gene of HL-60 cells which infected with Ad-NLS-RARα were both obviously higher than that of the cells which infected with Ad-KZ and non-infected (P<0.05). The proliferation ability of HL-60 cell infected with Ad-NLS-RARα was significantly increased (P<0.05). The level of CD11b of HL-60 cell infected with Ad-NLS-RARα and induced by ATRA was clearly decreased than control groups (P<0.05). The mRNA and protein levels of C-MYC gene of HL-60 cells infected with Ad-NLS-RARα and induced by ATRA were both obviously higher than that of the cells which infected with Ad-KZ and non-infected (P<0.05). Conclusion The recombined adenovirus Ad-NLS-RARα can increase the proliferation ability of HL-60 cell, and inhibit the differentiation of HL-60 cell through reduce the expression level of C-MYC gene.