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目的 :探讨三磷酸腺苷 (ATP)诱导 U93 7细胞凋亡过程中 p73 m RNA的表达变化 ,以进一步研究 p73基因在 U93 7细胞凋亡中的作用。方法 :用 ATP诱导 U93 7细胞凋亡 ,凋亡指标采用细胞形态学、DNA片段电泳、流式细胞术检测 DNA含量及细胞周期的变化等方法 ;采用半定量反转录 PCR(RT-PCR)检测 p73 m RNA的表达变化。结果 :ATP可诱导 U93 7细胞凋亡。 0 .2 5 g/L的ATP作用于 U93 7细胞 2 4h,光镜下见细胞出现较明显的聚集现象 ,胞膜皱缩 ,细胞体积缩小 ,Wright’ s+ Giemsa染色见胞核浓缩、核碎裂等现象 ,DNA片断电泳见清晰的梯形条带 ;流式细胞仪检测 :0 .2 5 g/L ATP作用于 U93 7细胞 2 4h、3 6h、48h,细胞的凋亡率分别为 2 .3 3 %、11.90 %、3 5 .49% ,并使细胞阻滞在 G2 ～ M+ S期 ,而对照组细胞凋亡率仅为 1.0 9% ;半定量 RT-PCR结果 :与对照组相比 ,仅 48h组 p73 m RNA的表达下调。结论 :ATP可诱导 U93 7细胞凋亡。在 U93 7细胞凋亡早期 ,p73 m RNA表达差异无显著性
Aims: To investigate the expression of p73 m RNA during the apoptosis of U93 7 cells induced by adenosine triphosphate (ATP) to further investigate the role of p73 gene in the apoptosis of U93 7 cells. Methods: Apoptosis of U937 cells was induced by ATP. The apoptosis index was determined by cell morphology, DNA fragment electrophoresis and flow cytometry. The semi-quantitative reverse transcription PCR (RT-PCR) Detection of p73 m RNA expression changes. Results: ATP induced the apoptosis of U93 7 cells. 0. 25 g / L of ATP on U93 7 cells for 2 4 h, under light microscopy, the cells showed more obvious aggregation phenomenon, cell membrane shrinkage, cell volume reduction, Wright ’s + Giemsa staining, nuclear condensation, nuclear broken The results of flow cytometry showed that the apoptotic rates of U93 7 cells were 24 h, 36 h and 48 h after treated with 0.25 g / L ATP respectively. 3 3%, 11.90%, 35.49% respectively, and the cells were arrested in G2 ~ M + S phase, while the apoptosis rate in the control group was only 1.09%. Semi-quantitative RT-PCR results: Compared with the control group , P73 m RNA expression was down-regulated only in 48h group. Conclusion: ATP can induce the apoptosis of U93 7 cells. At the early stage of U93 7 cell apoptosis, the expression of p73 m RNA showed no significant difference