外源性Rb基因导入对U937细胞周期和p21基因表达的影响

Effect of Exogenous Rb Gene on Expression of p21 and cell cycle regulation in U937 Cells

目的利用Rb基因重组腺病毒载体(AdCMV-Rb),将外源性Rb基因导入U937细胞,研究其对细胞周期、细胞凋亡及原癌基因p21表达的影响。方法AdCMV-Rb导入U937细胞后,用流式细胞术分析细胞周期和细胞凋亡率及Rb蛋白和p21蛋白的表达;RT-PCR检测原癌基因p21的表达。结果Ad-CMV-Rb可以高效导入U937细胞,Rb基因导入使U937细胞主要阻滞在G1期,细胞凋亡率在Rb基因导入24 h后无明显改变,48 h后凋亡率增高;Rb蛋白和p21蛋白表达增强;RT-PCR结果显示Rb基因的导入可以上调p21基因的表达。结论外源性Rb基因的导入可以上调原癌基因p21的表达,使得U937细胞周期阻滞在G1期。

Objective: To study the influence of Rb gene on the cell cycle, cell apoptosis and expression of p21 in U937 cells by recombinant adenovirus vector. Methods: Cell cycle and cell apoptosis, expression of Rb and p21 protein were detected by flow cytometric analysis. Expression of p21 gene were detected by RT-PCR. Results: Rb gene could be quickly and effectively transferred into the U937cells by adenovirus vector. Cell cycle was inhibited with most of the cells arrested in G1 phase. Apoptosis rate increased remarkably after AdCMV-Rb transferred for 48 h. Rb and p21 protein expression were up-regulated. Conclusion: AdCMV-Rb transfection can inhibit the cells cycle in G1 phase by up-regulating p21, it maybe has a potential significance on atherogenesis.