Regulation of pancreatic stellate cell activation by Notch3.
Regulation of pancreatic stellate cell activation by Notch3.
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DOI：
10.1186/s12885-017-3957-2
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发表时间：
2018-01-05
期刊：
影响因子：
3.8
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Song H;Zhang Y
活化的胰腺星状细胞（PaSC）是胰腺导管腺癌（PDAC）患者胰腺间质中癌相关成纤维细胞的关键细胞来源，但其活化机制尚不清楚。Notch信号传导途径，其组分在基质细胞中表达，参与包括肺和肝在内的几种器官的纤维化。在目前的研究中，我们研究了Notch信号转导是否参与PDAC中PaSC的激活。通过免疫组织化学染色检查人PDAC中Notch信号传导途径组分的表达，并使用RT-qPCR和蛋白质印迹法在小鼠PaSC中进行评估。使用免疫荧光、RT-qPCR和蛋白质印迹法评估PDAC基质细胞和活化小鼠PaSC中Notch 3的表达。采用RT-qPCR和western blotting检测siRNA介导的Notch 3敲低对PaSC活化的影响，采用CCK-8测定和划痕实验检测对PaSC增殖和迁移的影响。采用CCK-8法和划痕实验检测Notch 3 siRNA激活的PaSCs条件培养液对胰腺癌（LTPA）细胞的作用。使用Student t检验分析数据的统计学显著性。Notch 3在人PDAC基质细胞和活化的小鼠PaSC中过表达，并且用Notch 3 siRNA敲低Notch 3降低小鼠PaSC的增殖和迁移。与PaSC活化相关的标志物如α-平滑肌肌动蛋白（α-SMA）、胶原蛋白I和纤连蛋白的水平响应于Notch 3敲低而降低，表明Notch 3在PaSC活化中起重要作用。此外，我们证实，通过Notch 3 siRNA抑制PaSC活化减少了PaSC诱导的小鼠胰腺癌（LTPA）细胞的增殖和迁移。抑制PaSC中的Notch 3可以抑制PaSC的激活、增殖和迁移，并降低PaSC诱导的促肿瘤发生效应。因此，在PaSC中Notch 3沉默是PDAC患者的潜在新治疗选择。本文的在线版本（10.1186/s12885-017-3957-2）包含补充材料，可供授权用户使用。
Activated pancreatic stellate cells (PaSCs) are the key cellular source of cancer-associated fibroblasts in the pancreatic stroma of patients with pancreatic ductal adenocarcinoma (PDAC), however, the activation mechanism of PaSCs is not yet known. The Notch signaling pathway, components of which are expressed in stromal cells, is involved in the fibrosis of several organs, including the lung and liver. In the current study, we investigated whether Notch signal transduction is involved in PaSC activation in PDAC. The expression of Notch signaling pathway components in human PDAC was examined via immunohistochemical staining and assessed in mouse PaSCs using RT-qPCR and western blotting. Notch3 expression in both PDAC stromal cells and activated mouse PaSCs was evaluated using immunofluorescence, RT-qPCR and western blotting. The impact of siRNA-mediated Notch3 knockdown on PaSC activation was detected with RT-qPCR and western blotting, and the impact on PaSC proliferation and migration was detected using CCK-8 assays and scratch experiments. The effect of conditioned medium from PaSCs activated with Notch3 siRNA on pancreatic cancer (LTPA) cells was also detected with CCK-8 assays and scratch experiments. The data were analyzed for statistical significance using Student’s t-test. Notch3 was overexpressed in both human PDAC stromal cells and activated mouse PaSCs, and Notch3 knockdown with Notch3 siRNA decreased the proliferation and migration of mouse PaSCs. The levels of markers related to PaSC activation, such as α-smooth muscle actin (α-SMA), collagen I and fibronectin, decreased in response to Notch3 knockdown, indicating that Notch3 plays an important role in PaSC activation. Furthermore, we confirmed that inhibition of PaSC activation via Notch3 siRNA reduced the proliferation and migration of PaSC-induced mouse pancreatic cancer (LTPA) cells. Notch3 inhibition in PaSCs can inhibit the activation, proliferation and migration of PaSCs and reduce the PaSC-induced pro-tumorigenic effect. Therefore, Notch3 silencing in PaSCs is a potential novel therapeutic option for patients with PDAC. The online version of this article (10.1186/s12885-017-3957-2) contains supplementary material, which is available to authorized users.
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