| 姜涵毅,钱丹雯,王永生.MircoRNA-214对吉非替尼敏感及耐药细胞增殖和凋亡的影响[J].中国肿瘤,2015,24(9):785-791. |
| MircoRNA-214对吉非替尼敏感及耐药细胞增殖和凋亡的影响 |
| The Effect of microRNA-214 on Growth and Apoptosis of Gefitinib Sensitive and Resistant Cells |
| 投稿时间：2014-12-03 |
| DOI：10.11735/j.issn.1004-0242.2015.09.A016 |
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| 中文关键词: miR-214 非小细胞肺癌 吉非替尼 获得性耐药 PTEN |
| 英文关键词:miR-214 non-small cell lung cancer gefitinib acquired resistance PTEN |
| 基金项目:国家自然科学基金（81301882）；中央高校青年教师苗圃项目（20620140118） |
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| 中文摘要: |
| 摘 要：［目的］ 研究mircoRNA-214（miR-214）对PC9肺腺癌及吉非替尼耐药细胞（PC9/GR）增殖和凋亡的影响。［方法］ 在PC9细胞中转染miR-214模拟物及PC9/GR中转染miR-214抑制剂，使用定量逆转录PCR（qRT-PCR）检测其表达。MTT检测细胞转染miR-214模拟物或其抑制剂后的存活及增殖。在PC9细胞中顺时转染miR-214模拟物以检测上调miR-214对PC9细胞耐药性的影响，在PC9/GR细胞中顺时转染miR-214抑制物以检测下调miR-214对PC9/GR细胞耐药性的影响，并用流式细胞仪检测细胞的凋亡。Western blotting检测PTEN在PC9和PC9/GR细胞中的表达。构建PTEN 3’-UTR荧光素酶报告质粒验证miR-214的靶基因；建立异种移植模型检测miR-214抑制物对肺癌移植瘤的影响。［结果］ PC9细胞中miR-214低表达，上调miR-214的表达后PC9细胞对吉非替尼的敏感性降低，并且抵抗吉非替尼诱导的凋亡；而在PC9/GR细胞中低表达miR-214后，下调miR-214增加PC9/GR细胞对吉非替尼的敏感性，并且可以增强吉非替尼诱导的凋亡作用。荧光素酶报告载体实验证实PTEN是miR-214在细胞内的靶基因。动物异种移植模型表明miR-214抑制物可以增强PC9/GR对吉非替尼的敏感性。［结论］ MiR-214可能通过靶基因PTEN调控吉非替尼的获得性耐药。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the effect of microRNA-214 on the growth and apoptosis of Gefitinib sensitive and resistant cell lines.［Methods］ qRT-PCR was performed to detect the expression of miR-214 in PC9 and PC9/GR after transfection. MTT was used to analyze the survival and growth of cells. miR-214 mimic was simultaneously trasfected into PC9 cells and the cells response to miR-214 was analyzed；miR-214 inhibitor was transfected into PC9/GR and then investigate the cells’ sensitivity to Gefitinib. The apoptosis was detected by flow cytometry and the PTEN expression was detected by Western-blot. Dual luciferase system was establised to demonstrate the downstream target of miR-214 in tumor cells. Xenografte mice models were established to test the miR-214 impact in vivo.［Results］ The result showed that in Gefitinib sensitive cell line-PC9，miR-214 was down-regulated，and if miR-214 was up-regulated，PC9 became resistant to apoptosis induced by Gefitinib. On contrast，miR-214 was found up-regulated in PC9/GR (Gefitinib resistant cell line)，if the miR-214 was knock down，PC9/GR became more sensitive to Gefitinib. Dual luciferase report system demonstrated that PTEN was the downstream effector of miR-214 in tumor cells. Moreover，the xenograft models demonstrated that miR-214 inhibitor was able to induce the sensitivity of tumor cells to Gefitinib.［Conclusion］ miR-214 might regulate the Gefitinib acquired resistance via PTEN. |
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