Beclin1 inhibition promotes autophagy and decreases gemcitabine-induced apoptosis in Miapaca2 pancreatic cancer cells.
Beclin1 inhibition promotes autophagy and decreases gemcitabine-induced apoptosis in Miapaca2 pancreatic cancer cells.
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DOI：
10.1186/1475-2867-13-26
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发表时间：
2013-03-13
影响因子：
5.8
通讯作者：
Wang H
中科院分区：
文献类型：
--
作者：
Li X;Yan J;Wang L;Xiao F;Yang Y;Guo X;Wang H
Beclin 1是一个众所周知的自噬关键调节因子，也是一个单倍不足的肿瘤抑制因子。目前的研究表明，Beclin 1的下调或单等位基因缺失在各种癌症中经常被发现。本研究旨在探讨Beclin 1抑制剂对胰腺癌细胞自噬和吉西他滨诱导凋亡的影响。Beclin 1的表达被SiRNA转导抑制，并通过实时PCR和Western印迹法测定基因表达。Western blot检测Beclin 1抑制剂对Miapaca 2细胞自噬和吉西他滨诱导的细胞凋亡的影响。我们观察到，Beclin 1沉默促进微管相关蛋白1轻链3-II（LC 3-II）蛋白的形成，并在转染绿色荧光蛋白（GFP）标记的LC 3的Miapaca 2细胞中增加点状荧光信号。Beclin 1抑制剂对吉西他滨诱导的Miapaca 2细胞凋亡具有更强的抑制作用。我们的数据表明，Beclin 1沉默不仅上调自噬过程，而且在细胞凋亡的调节中发挥重要作用。Beclin 1抑制可抑制吉西他滨诱导的Miapaca 2细胞凋亡信号。
Beclin1 is a well-known key regulator of autophagy, which is also a haploinsufficient tumor suppressor. Current studies revealed that down-regulation or monoallelic deletions of Beclin1 were frequently found in various cancers. The purpose of this study was to investigate the effects of Beclin1 inhibition on autophagy and Gemcitabine-induced apoptosis of pancreatic cancer cells. Beclin1 expression was inhibited by siRNA transduction and gene expression was determined by Real-time PCR and Western blot. The effects of Beclin1 inhibition on autophagy and Gemcitabine-induced apoptosis of Miapaca2 cells were analyed through LC3 expression, cell viability, cell cycle and apoptosis by using Western blot. We observed that Beclin1 silence promoted microtubule-associated protein 1 light chain 3-II (LC3-II) protein formation and increased punctate fluorescent signals in Miapaca2 cells transfected with green fluorescent protein (GFP)-tagged LC3. Beclin1 inhibition showed a greater suppressive effect on Gemcitabine-induced apoptosis of Miapaca2 cells. Our data suggested that Beclin1 silence not only up-adjusted autophagy process, but also played an important role in the regulation of apoptosis. Beclin1 inhibition could inhibit apoptosis signaling induced by Gemcitabine in Miapaca2 cells.
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