| 肖国辉,厉娜,魏艳,徐会,彭晋,周福祥.辐射诱导线粒体功能障碍激活转化生长因子β1通路促进胰腺癌细胞上皮间质转化[J].中华放射医学与防护杂志,2021,41(6):407-412 |
| 辐射诱导线粒体功能障碍激活转化生长因子β1通路促进胰腺癌细胞上皮间质转化 |
| Mitochondrial dysfunction-induced expression of TGF-β1 pathway promoting epithelial-mesenchymal transition in pancreatic cancer cells after X-ray exposure |
| 投稿时间：2021-01-27 |
| DOI：10.3760/cma.j.issn.0254-5098.2021.06.002 |
| 中文关键词: X射线 线粒体 胰腺癌 迁移性 转化生长因子β1 |
| 英文关键词:X-rays Mitochondria Pancreatic cancer Migration TGF-β1 |
| 基金项目:国家自然科学基金（81903136） |
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| 摘要点击次数: 2558 |
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| 中文摘要: |
| 目的 研究X射线照射后胰腺癌细胞上皮间质转化的发生特点，探究线粒体功能障碍及其诱导的转化生长因子β1（TGF-β1）表达增加在上皮间质转化中的作用。方法 采用6 MV X射线多次（2 Gy×20次或4 Gy×10次）照射胰腺癌细胞株PATU1 988 t，利用transwell小室检测细胞迁移性，采用实时荧光定量方法检测上皮间质转化（EMT）相关因子E-cadherin、Vimentin、N-cadherin和MMPs家族（MMP2、MMP9）及线粒体复合体I的关键亚基及TGF-β1的转录水平变化。应用线粒体氧化磷酸化解偶联剂碳酰氰基-对-氯苯腙（CCCP）破坏线粒体膜电位，检测复合体亚基、TGF-β1含量及EMT相关分子的变化。应用小干扰RNA抑制TGF-β1的表达后，检测上皮间质转化及迁移变化。结果 多次照射后，存活肿瘤细胞迁移数增加（t=21.90、35.64，P<0.05），上皮间质转化增强，表现为上皮间质转化分子E-cadherin表达下降（t=8.37、6.77，P<0.05），N-cadherin （t=4.42、4.77，P<0.05）、Vimentin （t=4.57、3.02，P<0.05）、MMP2（t=7.27、26.08，P<0.05）和MMP9（t=13.26、7.29，P<0.05）表达均增加，TGF-β1（t=90.49、35.17，P<0.05）的含量也增加。沉默TGF-β1后细胞上皮间质转化及迁移性下降（t=38.66、11.54，P<0.05）。细胞发生线粒体功能障碍，具体表现为线粒体膜电位（t=6.94、29.71，P<0.05）和复合体相关亚基的表达量下降；加入CCCP破坏线粒体膜电位后（t=16.51，P<0.05），复合体亚基含量下降，TGF-β1的表达量增加（t=47.93，P<0.05），上皮间质转化增强。结论 辐射破坏了线粒体功能，进而激活了TGF-β1的表达，诱导胰腺癌细胞发生上皮间质转化，迁移性增强。 |
| 英文摘要: |
| Objective To investigate epithelial-mesenchymal transition and to explore the effects of mitochondrial dysfunction and increased expression of TGF-β1 pathway on epithelial-mesenchymal transition (EMT) in pancreatic adenocarcinoma after X-ray irradiation. Methods Split-dose irradiations of total 40 Gy (2 Gy×20 and 4 Gy×10) of 6 MV X-rays were performed on PATU1 988 t cells. The migration of the cells were examined through transwell filter chambers. Real-time PCR was adopted to detect the expression of EMT-related factors E-cadherin, Vimentin, N-cadherin, and MMPs (MMP2 and MMP9), critical subunits of mitochondrial complex I, and TGF-β1. The expression of EMT-related factors and content of TGF-β1 was detected after carbonylcyanide-m-chlorophenylhydrazone(CCCP) treatment. Meanwhile, the migration potential of pancreatic cells was detected after small interfering RNA (siRNA) knockdown of the expression of TGF-β1. Results After irradiation, the migration capacities of the cancer cells increased (t=21.90, 35.64, P<0.05). The expression of N-cadherin (t=4.42, 4.77, P<0.05), Vimentin (t=4.57, 3.02,P<0.05), MMP2 (t=7.27, 26.08, P<0.05), and MMP9 (t=13.26, 7.29, P<0.05) all increased, while the expression of E-cadherin deceased (t=8.37, 6.77, P<0.05). The expression of TGF-β1 (t=90.49, 35.17, P<0.05) increased. The expression of TGF-β1 decreased with small interfering RNA, which paralleled the inhibition of the epithelial-mesenchymal transition and migration (t=38.66, 11.54, P<0.05). Mitochondrial dysfunction was reflected by the decline in the membrane potential (t=6.94, 29.71, P<0.05) and complex-related subunits. The expression of TGF-β1 (t=47.93, P<0.05) and EMT-related factors further increased after mitochondrial function was destroyed (t=16.51,P<0.05). Conclusions Radiation-induced mitochondrial dysfunction can increase the expression of TGF-β1, which promotes epithelial-mesenchymal transition, and result in the migration of pancreatic cancer cell line. |
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