| PKM2基因沉默对人肺癌细胞生物学特性的影响 |
| 投稿时间：2017-04-16 修订日期：2017-05-31 点此下载全文 |
| 引用本文：张海红,陈虹宇,王亚南.PKM2基因沉默对人肺癌细胞生物学特性的影响[J].医学研究杂志,2018,47(9):63-67 |
| DOI：
10.11969/j.issn.1673-548X.2018.09.015 |
| 摘要点击次数: 1000 |
| 全文下载次数: 707 |
|
| 基金项目:中国医学科学院医学与健康科技创新工程项目（2016-I2M-1-002） |
|
| 中文摘要:目的 探讨M2型丙酮酸激酶（pyruvate kinase M2，PKM2）下调对人肺癌A549细胞生物学特性的影响。方法 采用免疫荧光技术检测A549细胞中PKM2蛋白的表达；将构建有PKM2-shRNA的重组质粒转入A549细胞；分别采用实时荧光定量PCR法和蛋白质印迹法检测PKM2 mRNA和蛋白表达水平；CCK-8法检测PKM2基因沉默对A549细胞增殖能力的影响；采用葡萄糖测定试剂盒测定细胞对葡萄糖的消耗量；Transwell实验检测细胞穿过人工基膜的能力。结果 肺癌A549细胞的细胞质及细胞核中均有PKM2表达；PKM2-shRNA质粒转染A549细胞48h后，可明显下调A549细胞中PKM2 mRNA及蛋白的表达水平，并且明显抑制A549细胞增殖；PKM2 shRNA可减少A549细胞葡萄糖的消耗，还可抑制细胞侵袭能力。结论 在肺癌细胞A549中，抑制PKM2表达可显著抑制细胞增殖、侵袭和糖代谢能力。 |
| 中文关键词:肺癌 M2型丙酮酸激酶 shRNA干扰 生物学功能 |
| |
| Influence of PKM2 Gene Silencing on Biological Characteristics of Lung Cancer Cells |
|
|
| Abstract:Objective To investigate the effect of pyruvate kinase M2 (PKM2) down-regulation on the biological characteristics of human lung cancer A549 cells. Methods The expression of PKM2 in A549 cells was detected by immunofluorescence. The recombinant plasmids of PKM2-shRNA were transferred into A549 cells. The expression levels of PKM2 mRNA and protein were test by real-time PCR and Western blotting respectively.CCK-8 was used to detect the effect of PKM 2 gene silencing on the proliferation of A549 cells. The glucose consumption was measured by glucose assay kit; Transwell test was used to detect the ability of cells to pass through the artificial basement membrane. Results The expression of PKM2 was found in cytoplasm and nucleus of lung cancer A549 cells. 48 hours after PKM2-shRNA plasmid transfection, PKM2 mRNA level and protein expression in A549 cells were significantly down-regulated, and the cell proliferation ability of A549 cells was significantly inhibited. PKM2 shRNA can reduce the consumption of glucose and invasion ability in A549 cells. Conclusion In lung cancer A549 cells, inhibition of PKM2 expression can significantly inhibit cell proliferation,metabolic capacity and invasion ability. |
| keywords:Lung cancer Pyruvate kinase M2 shRNA interference Biological function |
| 查看全文 查看/发表评论 下载PDF阅读器 |
