| 宋锐,袁金金,侯歌,等.lncRNA CCAT1靶向miR-130b-3p对人胰腺癌细胞PANC-1放射敏感性的影响[J].中华放射医学与防护杂志,2019,39(4):247-254.Song Rui,Yuan Jinjin,Hou Ge,et al.LncRNA CCAT1 enhances radiosensitivity of human pancreatic cancer cells PANC-1 by targeting miR-130b-3p[J].Chin J Radiol Med Prot,2019,39(4):247-254 |
| lncRNA CCAT1靶向miR-130b-3p对人胰腺癌细胞PANC-1放射敏感性的影响 |
| LncRNA CCAT1 enhances radiosensitivity of human pancreatic cancer cells PANC-1 by targeting miR-130b-3p |
| 投稿时间：2018-09-05 |
| DOI：10.3760/cma.j.issn.0254-5098.2019.04.002 |
| 中文关键词: lncRNA CCAT1 miR-130b-3p 胰腺癌 放射敏感性 |
| 英文关键词:lncRNA CCAT1 miR-130b-3p Pancreatic cancer Radiosensitivity |
| 基金项目:国家重点研发计划项目（2016YFC0105713） |
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| 中文摘要: |
| 目的 探讨lncRNA CCAT1和miR-130b-3p对体外培养的胰腺癌细胞PANC-1放射敏感性的影响。方法 采用Real-time PCR检测胰腺癌组织及其细胞系和2 Gy X射线照射后PANC-1细胞中CCAT1和miR-130b-3p的相对表达水平。沉默CCAT1表达、抑制miR-130b-3p表达后，应用流式细胞仪、Caspase 3活性检测试剂盒及克隆形成实验检测细胞凋亡率、Caspase 3活性和细胞存活分数，并绘制单击多靶模型拟合曲线；利用starBase v2.0在线预测、荧光素酶报告基因、RNA结合蛋白免疫沉淀实验（RIP）及Real-time PCR实验，验证CCAT1和miR-130b-3p的靶向关系。结果 在放射抵抗的胰腺癌组织、胰腺癌细胞系和2 Gy照射的PANC-1细胞中，CCAT1表达均上调（t=6.322~8.555，P<0.05），miR-130b-3p表达下调（t=3.950~18.795，P<0.05）。2 Gy照射并沉默CCAT1，PANC-1细胞存活分数降低（t=2.929、5.047、5.234、5.125，P<0.05），细胞凋亡率增加（t=6.953，P<0.05），Caspase 3活性升高（t=6.836，P<0.05）。发现CCAT1能靶向调控miR-130b-3p表达，抑制miR-130b-3p表达，PANC-1细胞存活分数增大（t=4.564、6.736、8.656，P<0.05），细胞凋亡减少（t=5.234，P<0.05），Caspase 3活性降低（t=10.440，P<0.05）。结论 沉默CCAT1表达能够促进miR-130b-3p表达，从而增加PANC-1细胞放射敏感性。 |
| 英文摘要: |
| Objective To investigate the effect of lncRNA CCAT1 and miR-130b-3p on the radiosensitivity of human pancreatic cancer cells PANC-1. Methods Real-time PCR was used to detect the relative expression levels of CCAT1 and miR-130b-3p in pancreatic cancer tissues and cell lines including PANC-1 cells irradiated with 2 Gy X-rays. After silencing CCAT1 and/or inhibiting miR-130b-3p expression, cell apoptosis rate, Caspase 3 activity and cell survival were detected by flow cytometry, Caspase 3 activity detection kit and colony formation assay, respectively. Cell survival curve was stimulated by the multi-target single-hit model. Based on the starBase v2.0 online analysis, the luciferase reporter gene assay, RNA-binding protein immunoprecipitation assay (RIP) and Real-time PCR assay were applied to verify the relationship between CCAT1 and miR-130b-3p. Results CCAT1 expression was up-regulated (t=6.322-8.555, P<0.05), but miR-130b-3p expression was down-regulated (t=3.950-18.795, P<0.05) in the radiation-resistant pancreatic cancer tissues, pancreatic cancer cell lines and 2 Gy-irradiated PANC-1 cells. When the CCAT1 silenced PANC-1 cells were irradiated with 2 Gy, cell survival fraction decreased (t=2.929, 5.047, 5.234, 5.125, P<0.05), apoptosis rate and Caspase 3 activity increased (t=6.953, 6.836, P<0.05). CCAT1 could selectively regulate miR-130b-3p expression. Inhibition of miR-130b-3p expression could enhance PANC-1 cell survival (t=4.564, 6.736, 8.656, P<0.05), but reduced apoptosis rate (t=5.234, P<0.05) and Caspase 3 activity (t=10.440, P<0.05). Conclusions Silencing CCAT1 promotes the expression of miR-130b-3p and enhances radiosensitivity of PANC-1 cells. |
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