| 张巍 韩磊 光洪德 刘国东 贺梁 周文平.癌基因Src 在骨肉瘤细胞侵袭伪足形成中的作用[J].,2015,15(14):2633-2636 |
| 癌基因Src 在骨肉瘤细胞侵袭伪足形成中的作用 |
| Effects of Oncogene Src on Invadopodia Formation of Osteosarcoma Cells |
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| DOI： |
| 中文关键词: 骨肉瘤 侵袭伪足 侵袭、转移 Src基因 |
| 英文关键词: Osteosarcoma cells Invadopodia Invasion and Metastasis Src gene |
| 基金项目:国家自然科学基金青年基金项目（31401215）；辽宁省科技攻关计划项目（2013225220） |
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| 中文摘要: |
| 目的：探讨癌基因Src在体外培养骨肉瘤细胞侵袭伪足形成中的作用。方法：构建Src shRNA慢病毒表达载体，在HEK293T细
胞中包装慢病毒，感染HT-1080 骨肉瘤细胞，经嘌呤霉素加压筛选，获得稳定沉默Src 基因的骨肉瘤细胞系HT-1080-shSrc；实时
定量PCR和Western Blot法检测基因沉默效率；采用原位明胶酶谱法检测侵袭伪足形成；采用侵袭小室实验检测下调Src基因表
达对HT-1080 细胞侵袭力的影响。结果：成功构建稳定沉默Src基因的骨肉瘤细胞系HT-1080-shSrc及对照细胞系HT-1080-shluc，
经实时定量PCR和Western Blot 检测，与对照细胞系相比，HT-1080-shSrc 细胞中Src 基因表达下调3 倍以上；下调HT-1080 细
胞中Src 基因表达能显著抑制HT-1080 细胞侵袭伪足形成及其对细胞外基质的降解能力；下调Src 基因表达能显著抑制骨肉瘤
细胞侵袭力。结论：癌基因Src 参与调节骨肉瘤细胞HT-1080 侵袭伪足形成，促进肿瘤侵袭、转移。 |
| 英文摘要: |
| Objective:To investigate the effects of oncogene Src on invadopodia formation of osteosarcoma cells (HT-1080) in
vitro.Methods:Src shRNA lentiviral vector was constructed, packaged in HEK293T cells. And HT-1080 cells were infected, then
screened by puromycin. And then stablely Src gene silencing osteosarcoma cell line HT-1080-shSrc was obtained. Quantification PCR
and Western Blot were carried out to detect the gene silencing efficiency. Invadopodia formation was observed by in situ gelatin
zymography. In vitro invasion assays were used to detect the effects of downregulation Src expression on invasive ability of HT-1080
cells.Results:Stablely Src gene silencing osteosarcoma cell line HT-1080-shSrc and control cell line HT-1080-shluc were successfully
constructed. Quantification PCR and Western Blot showed, compared with HT-1080-shluc cells, the expression of Src in HT-1080-shSrc
cells decreased more than 3 times. Invadopodia formation, the ability to degradate the extracellular matrix and the invasive ability were
significantly inhibited when Src expression was downregulated in HT-1080 cells.Conclusion:Oncogene Src is involved in regulating
invadopodia formation in osteosarcoma cells HT-1080, and promoting tumor invasion and metastasis. |
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