| miR-152通过靶向FGF2抑制肺癌A549细胞系的增殖侵袭 |
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| 引用本文： | 何磊,杜佳辉,李娜.miR-152通过靶向FGF2抑制肺癌A549细胞系的增殖侵袭[J].天津医科大学学报,2022,0(2):145-150,185. |
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| 作者姓名： | 何磊 杜佳辉 李娜 |
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| 作者单位： | （1.郑州人民医院急诊科，郑州 450000；2.河南省胸科医院胸外科，郑州450000） |
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| 摘 要： | 目的:探讨miR-152通过靶向成纤维细胞生长因子2（FGF2）对肺癌A549细胞系的增殖和侵袭的作用机制。方法:通过实时荧光定量PCR（qPCR）检测正常上皮细胞BEAS-2B和肺癌A549细胞中miR-152和FGF2的表达情况；采用转染技术，分别特异性表达肺癌A549细胞系中miR-152和FGF2基因的表达。将肺癌A549细胞系分为6组:miRNA 空转组（miR-NC组）、miR-152 过表达组（miR-152 mimics组）、miR-152 抑制组（miR-152 inhibitor组）、FGF2过表达空转组（pcDNA3.1-NC组）、FGF2过表达组（pcDNA3.1-FGF2组）和miR-152过表达+FGF2过表达组（miR-152+ pcDNA3.1-FGF2组）。通过qPCR检测A549细胞系中miR-152和FGF2的表达水平；细胞克隆和Transwell小室实验分别检测A549细胞系的增殖和侵袭能力；采用双荧光报告基因法验证miR-152与FGF2结合情况；Western印迹法检测A549细胞系中FGF2、自噬标志蛋白微管相关蛋白1轻链3B（LC3B）Ⅱ/Ⅰ、beclin1和p62的表达量。结果:qPCR结果显示，与正常上皮细胞BEAS-2B比较，肺癌A549细胞中miR-152的表达量显著降低（P＜0.01），而FGF2的表达量显著增加（P＜0.01）。TargetScan生物信息学软件预测显示miR-152与FGF2的3′UTR结合，双荧光报告基因检测结果显示，miR-152与FGF2直接结合。细胞克隆和Transwell小室实验结果显示，与miR-NC组比较，miR-152 mimics组中的A549细胞系的增殖和侵袭能力显著减低（P＜0.05），miR-152 inhibitor组的增殖和侵袭能力显著增加（P＜0.05）。与pcDNA3.1-NC组比较，pcDNA3.1-FGF2组中的A549细胞系的增殖和侵袭能力显著增加（P＜0.05）。与miR-152 mimics组比较，miR-152+ pcDNA3.1-FGF2组中A549细胞系的增殖和侵袭能力显著增加（P＜0.05）。Western印迹结果显示，与miR-NC组比较，miR-152 mimic组FGF2和p62蛋白表达显著降低（P＜0.05），但LC3B Ⅱ/Ⅰ和beclin1的表达量显著增加（P＜0.05），而miR-152 inhibitor组结果与之相反。与pcDNA3.1-NC组比较，pcDNA3.1-FGF2组FGF2和p62蛋白表达显著增加（P＜0.05），而LC3B Ⅱ/Ⅰ和beclin1的表达量显著降低（P＜0.05）。与miR-152 mimics组比较，miR-152+ pcDNA3.1-FGF2组FGF2和p62蛋白表达显著增加（P＜0.05），LC3B Ⅱ/Ⅰ和beclin1蛋白表达量显著降低（P＜0.05）。结论:miR-152靶向FGF2抑制肺癌A549细胞系自噬，从而降低增殖侵袭能力，可成为肺癌临床治疗的新靶点。
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| 关 键 词： | 肺癌 miR-152 成纤维细胞生长因子2 自噬 细胞增殖 细胞侵袭 |
The effect of miR-152 on the proliferation and invasion of lung cancer A549 cell line regulated by FGF2 |
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| HE Lei,DU Jia-hui,LI Na.The effect of miR-152 on the proliferation and invasion of lung cancer A549 cell line regulated by FGF2[J].Journal of Tianjin Medical University,2022,0(2):145-150,185. |
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| Authors: | HE Lei DU Jia-hui LI Na |
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| Affiliation: | （1.Department of Emergency，Zhengzhou People's Hospital，Zhengzhou 450000，China；2.Department of Thoracic Surgery，Henan Thoracic Hospital，Zhengzhou 450000，China） |
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| Abstract: | Objective: To investigate the mechanism of miR-152 targeting fibroblast growth factor 2（FGF2） on the proliferation and invasion of lung cancer A549 cell line. Methods: Real-time fluorescence quantitative PCR（qPCR） was used to detect the expression of miR-152 and FGF2 in normal epithelial cells BEAS-2B and lung cancer A549 cells. Transfection technology was used to specifically overexpress miR-152 and FGF2 in lung cancer A549 cell lines，respectively. The experiment divided the lung cancer A549 cell lines into 6 groups: miRNA idling group（miR-NC），miR-152 overexpression group（miR-152 mimics），miR-152 inhibitor group（miR-152 inhibitor），FGF2 overexpression idling group（pcDNA3.1-NC），FGF2 overexpression group（pcDNA3.1-FGF2） and miR-152 overexpression+FGF2 overexpression group（miR-152+ pcDNA3.1-FGF2）. qPCR was used to detect the expression levels of miR-152 and FGF2 in A549 cell line. Cell cloning and Transwell cell experiment were used to detect the proliferation and invasion ability of A549 cell line. Dual fluorescence reporter gene method was used to verify the binding of miR-152 and FGF2. Western blotting was used to detect the protein expression of FGF2，autophagy marker protein microtubule-associated protein 1 light chain 3B（LC3B） Ⅱ/Ⅰ，beclin1 and p62 in A549 cell line. Results: The qPCR results showed that compared with normal epithelial cells BEAS-2B，the expression of miR-152 in lung cancer A549 cells was significantly reduced（P＜0.01），while the expression of FGF2 was significantly increased（P＜0.01）. TargetScan bioinformatics software predicted that miR-152 was bound to the 3′UTR of FGF2，and dual-fluorescence reporter gene test results showed that miR-152 was directly bound to FGF2. The results of cell cloning and Transwell chamber experiments showed that compared with the miR-NC group，the proliferation and invasion ability of the A549 cell line in the miR-152 mimics group was significantly reduced（P＜0.05），and the proliferation and invasion of miR-152 inhibitor group increased significantly（P < 0.05）. Compared with the pcDNA3.1-NC group，the proliferation and invasion ability of the A549 cell line in the pcDNA3.1-FGF2 group increased significantly（P＜0.05）. Compared with miR-152 mimics group，the proliferation and invasion ability of A549 cell line in miR-152+pcDNA3.1-FGF2 group increased significantly（P＜0.05）. Western blotting results showed that，compared with the miR-NC group，the expression of FGF2 and p62 protein in the miR-152 mimic group was significantly reduced（P＜0.05），but the expression of LC3B Ⅱ/Ⅰ and beclin1 was significantly increased（P＜0.05），while the miR-152 inhibitor group result was the opposite. Compared with the pcDNA3.1-NC group，the expression of FGF2 and p62 protein in the pcDNA3.1-FGF2 group increased significantly（P＜0.05），while the expression of LC3B Ⅱ/Ⅰ and beclin1 decreased significantly（P＜0.05）. Compared with the miR-152 mimics group，the expression of FGF2 and p62 protein in the miR-152+pcDNA3.1-FGF2 group was significantly increased（P＜0.05），and the protein expression of LC3B Ⅱ/Ⅰ and beclin1 was significantly decreased（P＜0.05）. Conclusion: MiR-152 targeting FGF2 can inhibit autophagy of lung cancer A549 cell line，and may become a new target for clinical treatment of lung cancer. |
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| Keywords: | lung cancer miR-152 fibroblast growth factor 2 autophagy cell proliferation cell invasion |
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