目的：研究P53正向凋亡调节因子基因(P53 upregulate modulator of apoptosis,PUMA)对胰腺癌细胞株BxPC3凋亡的影响及其可能的作用机制。方法：以100 MOI的携PUMA基因重组腺病毒（AdPUMA）感染BxPC3细胞0～96 h，流式细胞术检测BxPC3细胞凋亡率，Western blotting检测BxPC3细胞中PUMA、Bcl2、Bax、Cytochrome C和Caspase3蛋白的表达，Western blotting检测BxPC3细胞中细胞质和线粒体内Bax的表达及Bax寡聚体。结果：随着AdPUMA感染时间的延长，BxPC3细胞凋亡率逐渐增加，48 h时最高。AdPUMA感染促进BxPC3细胞中PUMA、Cytochrome C和Caspase3 蛋白的表达，抑制BxPC3细胞中Bcl2蛋白的表达。AdPUMA感染后BxPC3细胞的凋亡率与BxPC3细胞中PUMA蛋白的表达具有明显的相关性。AdPUMA感染不影响BxPC3细胞中Bax蛋白的总表达量，但细胞质中的Bax几乎完全消失，而线粒体中的Bax表达明显增加；AdPUMA感染诱导BxPC3细胞中Bax蛋白的寡聚化。结论：PUMA基因通过线粒体途径促进胰腺癌细胞凋亡。

Objective:To investigate the effect of P53 upregulate modulator of apoptosis (PUMA) on the apoptosis of pancreatic carcinoma BxPC3 cells and the possible mechanism. Methods: BxPC3 cells were infected with recombinant adenovirus containing PUMA gene (AdPUMA) at 100 MOI for 096 h. Apoptosis of BxPC3 cells was examined by FCM. Expressions of PUMA, Bcl2, Bax, Cytochrome C and Caspase3 proteins in BxPC3 cells were detected by Western blotting. Bax expression in the cytoplasm and mitochondrion and Bax oligomer expression expression in BxPC3 cells were determined by Western blotting. Results:Apoptosis rates of BxPC3 cells were significantly increased with the time of AdPUMA infection, and peaked after 48 h. AdPUMA infection increased the expressions of PUMA, Cytochrome C and Caspase3 proteins in BxPC3 cells, and decreased the expression of Bcl2 protein. Apoptosis rate of BxPC3 cells after AdPUMA infection was correlated with PUMA expression. AdPUMA did not affect the expression of total Bax protein in BxPC3 cells, but Bax expression in cytoplasm was dramatically decreased after infection, and Bax expression in mitochondrion was markedly increased. Furthermore, AdPUMA infection induced Bax oligomerization in BxPC3 cells.Conclusion: PUMA can promote apoptosis of pancreatic carcinoma cells through mitochondrion pathway.

卫生部科学研究基金项目（No. WKJ200601010）