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目的:研究野生型p53基因重组腺病毒载体(AdCMV-p53)导入对U937细胞分化、凋亡和清道夫受体CD36表达的影响。方法:AdCMV-p53导入U937细胞后,用细胞计数、细胞周期分析、台盼蓝染色排除法计数细胞悬液中的活细胞数目和NBT还原反应观察其对U937细胞生长、分化的影响;RT-PCR、免疫荧光和流式细胞分析检测AdCMV-p53导入对CD36表达的影响。结果:AdCMV-p53可以高效导入U937细胞,野生型p53基因导入促进U937细胞向巨噬细胞分化,台盼蓝染色发现实验组阳性细胞数(64.6±9.2) %较对照组(14.2±5.5) %明显增多,吞噬能力增强;NBT还原反应实验组(49.7±12.6) %较对照组(6.3±1.8) %升高。RT-PCR和流式细胞分析检测,野生型p53基因导入使得CD36 mRNA转录增强,CD36蛋白表达增加。结论:野生型p53基因能影响细胞分化和凋亡,并上调清道夫受体CD36的表达,对于动脉粥样硬化的预防和基因治疗具有潜在意义。
Objective: To investigate the effects of AdCMV-p53 wild-type p53 gene on the differentiation and apoptosis of screened U937 cells and the expression of scavenger receptor CD36. Methods: U937 cells were transfected with AdCMV-p53. The number of viable cells and the NBT reduction reaction in U937 cells were counted by cell counting, cell cycle analysis and trypan blue exclusion method to observe the effect of AdCMV-p53 on the growth and differentiation of U937 cells. RT- PCR, immunofluorescence and flow cytometry analysis of AdCMV-p53 on CD36 expression. Results: AdCMV-p53 was transfected into U937 cells efficiently. Wild type p53 gene was introduced into U937 cells to promote the differentiation of U937 cells into macrophages. The number of positive cells (64.6 ± 9.2%) in experimental group was 14.2 ± 5.5% (49.7 ± 12.6)% in NBT reduction group was significantly higher than that in control group (6.3 ± 1.8)%. RT-PCR and flow cytometry analysis, wild-type p53 gene transfection led to increased CD36 mRNA transcription, CD36 protein expression increased. Conclusion: The wild-type p53 gene can affect cell differentiation and apoptosis, and up-regulate the expression of scavenger receptor CD36, which has potential significance for the prevention and gene therapy of atherosclerosis.