Impact of oxidative stress on the cytoskeleton of pancreatic epithelial cells
Impact of oxidative stress on the cytoskeleton of pancreatic epithelial cells
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DOI：
10.3892/etm.2014.1979
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发表时间：
2014-11-01
影响因子：
2.7
通讯作者：
Lu, Qi-Ping
中科院分区：
文献类型：
--
作者：
Hu, Wei-Guo;Lu, Qi-Ping
在本研究中，研究了活性氧对三维培养系统中胰腺上皮细胞形态变化的影响。此外，还测定了该过程中信号分子的表达。使用Matrigel(TM)构建胰腺上皮细胞和癌细胞的三维培养模型。用1或200μmol/l H2O2（一种典型的活性氧）刺激培养的细胞，然后在15分钟、1小时和4小时后评估形态变化。使用激光扫描共聚焦显微镜和免疫荧光染色观察细胞的细胞骨架。此外，使用ELISA检测活化B细胞的核因子κ轻链增强子（NF-κB）的核含量。结果表明，200 μmol/l H2O2处理15 min后细胞出现收缩，1 h后细胞形态恢复；然而，4小时后细胞尺寸减小。因此，H2O2 处理后细胞内肌动蛋白和微管迅速丢失，细胞骨架变得模糊，最终在 4 小时后解体。对于正常胰腺上皮细胞和癌细胞也观察到类似的结果。相比之下，用1μmol/l H2O2处理，不影响胰腺上皮细胞的形态和细胞骨架。此外，200 μmol/l H2O2处理逐渐增加NF-kappa B活性，而1 μmol/l H2O2处理对NF-kappa B活性影响不大。因此，证明氧化应激可诱导胰腺上皮细胞早期发生可逆性细胞收缩和细胞骨架去极化，并可增加NF-kappa B表达。
In the present study the effect of reactive oxygen species on the morphological changes of pancreatic epithelial cells in a three-dimensional culture system was investigated. In addition, the expression of signaling molecules during this process was determined. Matrigel (TM) was used to construct a three-dimensional culture model of pancreatic epithelial and cancer cells. The cultured cells were stimulated with 1 or 200 mu mol/l H2O2 (a typical reactive oxygen species), and the morphological changes were then evaluated after 15 min, 1 h and 4 h. The cytoskeleton of the cells was observed using laser scanning confocal microscopy with immunofluorescence staining. In addition, the nuclear content of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappa B) was detected using ELISA. The results demonstrated that treatment with 200 mu mol/l H2O2, induced cell contraction after 15 min, and cell morphology recovered after 1 h; however, cell size was reduced after 4 h. Consequently, intracellular actin and microtubules were rapidly lost following H2O2 treatment, and the cytoskeleton became indistinct and eventually disintegrated after 4 h. Similar observations were noted for the normal pancreatic epithelial and cancer cells. By contrast, treatment with 1 mu mol/l H2O2, did not affect the morphology and cytoskeleton of pancreatic epithelial cells. In addition, 200 mu mol/l H2O2 treatment increased the activity of NF-kappa B gradually, while 1 mu mol/l H2O2 treatment was found to have little impact on the activity of NF-kappa B. Therefore, it was demonstrated that oxidative stress can induce the early onset of reversible cell contraction and cytoskeleton depolarization in pancreatic epithelial cells, and can increase NF-kappa B expression.