| 董 熠,李营歌,姚 颐.人肺癌相关成纤维细胞的原代培养及其放射激活初探[J].肿瘤学杂志,2018,24(11):1071-7075. |
| 人肺癌相关成纤维细胞的原代培养及其放射激活初探 |
| Activation of Human Lung Cancer-associated Fibroblasts Induced by Irradiation |
| 投稿时间：2018-08-15 |
| DOI：10.11735/j.issn.1671-170X.2018.11.B007 |
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| 中文关键词: 人 癌相关成纤维细胞 原代培养 放射 活化 |
| 英文关键词:human cancer-associated fibroblasts primary culture radiation activation |
| 基金项目:国家自然科学基金（81670123，3080190，81372441） |
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| 摘要点击次数: 1815 |
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| 中文摘要: |
| 摘 要：［目的］ 探讨在体外培养条件下，不同剂量的X线照射对人肺癌相关成纤维细胞（cancer-associated fibroblasts，CAFs）表型的影响。［方法］ 用经组织块贴壁法从手术标本中提取人肺癌CAFs并原代培养。使用0Gy、4Gy、8Gy、12Gy、16Gy的不同剂量X线照射CAFs，通过Western blot、qRT-PCR检测其平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、波形蛋白（Vimentin）、成纤维细胞活化蛋白（fibroblast activation protein，FAP）、平足蛋白（podoplanin，PDPN）的蛋白和RNA水平；使用CCK8、划痕实验及流式细胞术检测细胞增殖和迁移及凋亡情况。［结果］ 与其他组比较，8Gy照射组的人肺CAFs中α-SMA、Vimentin、FAP、PDPN在蛋白和RNA水平表达更高。划痕实验提示8Gy照射组在48h首先愈合。CCK8研究提示该组细胞增殖活性明显高于其他照射组；流式细胞术显示照射后该组细胞凋亡率为37%，与0Gy组（36%）接近，明显低于其他照射剂量组。［结论］ 经过一定剂量的X线照射后，人肺CAFs可表达更高水平的标志蛋白、同时表现出更强的生长和迁移能力，而该表型变化在8Gy照射组最为显著，为将来深入探讨CAFs的放射激活机制奠定基础。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effect of irradiation on the activation of human lung cancer-associated fibroblasts (CAFs) in vitro. ［Methods］ The primarily cultured lung cancer-associated fibroblasts were irradiated with different dosage of X-ray (0，4，8，12 and 16Gy). The protein and mRNA expressions of α-SMA，vimentin，FAP and PDPN were detected by Weston blot and qRT-PCR，respectively. The proliferation and the migration ability of CAFs were determined by CCK8，wound healing and flow cytometry. ［Results］ Compared with other groups，the expression levels of α-SMA，vimentin，FAP，PDPN proteins and mRNA were significantly higher，and the migration ability was also significantly higher in the 8Gy. The cell proliferation was higher and apoptosis rate was lower in 8Gy group than those of other groups. ［Conclusion］ The irradiation of X-ray at dosage of 8Gy can up-regulate the expression of fibroblast molecular markers and enhance cell proliferation and migration，as well as inhibit cell apoptosis of human lung cancer-associated fibroblasts in vitro，indicating that the irradiation at appropriate dosage can induce the activation of CAFs. |
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