Partial Beclin 1 silencing aggravates doxorubicin-and Fasinduced apoptosis in HepG2 cells

瞄准：在 HepG2 房间的危险性上调查 Beclin 1 的角色在反船边交货抗体或 doxorubicin 治疗以后经历 apoptosis。方法：Beclin 1 silencing 用 RNA 干扰被完成。DNA 倍性， apoptotic 房间的百分比和 mitochondrial 膜电位被流动血细胞计数估计。多形核白细胞(自动数据处理核糖) 的 Beclin 1， Bcl-X (L) 和细胞色素 c，和劈开聚合酶(PARP ) 的层次是由使用西方的污点的 assayed。结果：Beclin 1 表示在 Beclin 1 siRNA transfection 以后由 75% 72 h 减少了。部分 Beclin 1 silencing 显著地增加了 subG1 房间的百分比有 doxorubicin 或反船边交货抗体的 24 和 40 h 术后疗法，分别地并且这 potentiation 被处理与一个 pan-caspase 禁止者废除。部分 Beclin 1 silencing 也增加了 PARP 劈开， mitochondrial 膜去极和 cytosolic 细胞色素 c。部分 Beclin 1 silencing 的 pro-apoptotic 后果没在 Bcl-X (L) 表示与衰落被联系。结论：部分 Beclin 1 silencing 在与反船边交货抗体或与 doxorubicin 对待的 HepG2 房间加重 mitochondrial permeabilization 和 apoptosis。

AIM： To investigate the role of Beclin 1 on the susceptibility of HepG2 cells to undergo apoptosis after anti-Fas antibody or doxorubicin treatment. METHODS： Beclin 1 silencing was achieved using RNA interference. DNA ploidy, the percentage of apoptotic cells and the mitochondrial membrane potential were assessed by flow cytometry. Levels of Beclin 1, BCI-XL and cytochrome c, and the cleavage of poly （ADP-ribose） polymerase （PARP） were assayed by using Western blots. RESULTS： Beclin 1 expression decreased by 75% 72 h after Beclin 1 siRNA transfection. Partial Beclin 1 silencing significantly increased the percentage of subG1 cells 24 and 40 h after treatment with doxorubicin or anti-Fas antibody, respectively, and this potentiation was abrogated by treatment with a pan-caspase inhibitor. Partial Beclin 1 silencing also increased PARP cleavage, mitochondrial membrane depolarization and cytosolic cytochrome c. The pro-apoptotic consequences of partial Beclin 1 silencing were not associated with a decline in Bcl-XL expression. CONCLUSION： Partial Beclin 1 silencing aggravates mitochondrial permeabilization and apoptosis in HepG2 cells treated with an anti-Fas antibody or with doxorubicin.