哇巴因诱导Jurkat T淋巴细胞凋亡及Bcl-2、Bax表达的变化

Apoptosis and the expression of Bcl-2 and Bax in Jurkat T lymphocytes induced by ouabain

目的探讨哇巴因对白血病细胞株Jurkat是否具有凋亡诱导作用,并进一步研究bcl2基因家族在此过程中的作用。方法四甲基偶氮唑盐(MTT)法观察哇巴因对Jurkat细胞生长的抑制程度;脱氧核苷酸末端转移酶(TdT)介导的缺口末端标记法等观察细胞凋亡;半定量逆转录PCR检测bcl2、bax基因表达水平;应用Westernblot检测Bcl2、Bax蛋白表达水平。结果哇巴因能诱导Jurkat细胞凋亡,细胞凋亡的数量与哇巴因的浓度和作用时间呈正相关,在此过程中出现bax转录,Bax蛋白表达增加。结论bcl2基因家族参与了哇巴因诱导Jurkat细胞凋亡的基因调控。

Objective To observe whether ouabain can induce the apoptosis of Jurkat cells and the role of bcl-2 gene family in this process. Methods Using the MTT method, the inhibitory effect of onabain on the proliferation was assayed. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end- labeling reaction method （TUNEL） was performed to observe apoptosis. The expression levels of bcl-2 and bax mRNA were detected by semi-quantitative reverse transcription PCR （RT-PCR） technique. Western blot analysis was applied to detect the protein expression of Bax and Bcl-2. Resalts Ouabain could induce apoptosis of Jurkat cells. The rate of apoptosis was correlated with the concentration of ouabain and treating time. The protein and mRNA expression of P, ax was increased after ouabain treatment. Conclusion Bcl-2 gene family may participate in the ouabain induced apoptosis of Jurkat cells.