| CXCR4基因沉默对黑色素瘤侵袭和转移影响的实验研究 |
| 投稿时间：2015-11-19 修订日期：2015-11-28 点此下载全文 |
| 引用本文：吴包金,李文鹏,丁伟,周兆平,周仲文,顾小叶,江华.CXCR4基因沉默对黑色素瘤侵袭和转移影响的实验研究[J].医学研究杂志,2016,45(5):53-58 |
| DOI：
10.11969/j.issn.1673-548X.2016.05.013 |
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| 基金项目:上海市卫生局科研课题(20114211) |
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| 中文摘要:目的 探讨CXCR4基因沉默对黑色素瘤侵袭及转移的影响及可能机制。方法 设计合成CXCR4特异性短发夹状RNA(shRNA)插入pSilencer载体,转染人高转移性黑色素瘤细胞株MV3、RT-PCR和Western blot法检测shRNA对靶基因CXCR4表达的影响。利用Transwell小室模型检测细胞体外侵袭能力,MTT法检测细胞体外增殖能力。通过裸鼠尾静脉瘤细胞注射,将转染后的细胞接种至裸鼠皮下,构建黑色素瘤转移模型,观察肿瘤生长情况。结果 CXCR4-shRNA能有效下调CXCR4基因的表达,降低黑色素瘤细胞增殖、体外侵袭及转移能力。RT-PCR及细胞免疫荧光显示,转染CXCR4-shRNA的黑色素瘤瘤MV3细胞系MMP-2表达下调;免疫组化结果显示,实验组裸鼠黑色素瘤瘤体及转移灶组织MMP-2的表达低于对照组。结论 shRNA介导的CXCR4基因沉默在体外实验能够显著抑制黑色素瘤细胞的生长和增殖活性,减弱体外侵袭能力,且体外产生抑瘤效应,明显降低肝、脑器官转移能力和定向肺转移能力。其机制可能与SDF-1/CXCR4信号途径阻断,进而抑制MMP-2的表达有关。 |
| 中文关键词:黑色素瘤 趋化因子受体CXCR4 基因沉默 肿瘤转移 |
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| Experimental Study on the Invasion and Metastasis of Melanoma by CXCR4 Gene Silence |
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| Abstract:Objective To study the effects on the invasion and metastasis of melanoma by CXCR4 gene silence in nude mice. Methods A CXCR4 specific recombinant plasmid vector was constructed and transfected into the cultured MV3 cell line with lipofectamine. RT-PCR and Western blot were used to detect the mRNA and protein expression of CXCR4. Invasion capability in vitro of the cells was evaluated by Transwell chamber. The cell proliferation capacity was detected by MTT method. The nude mouse model of metastasis was established by injection of MV3 cells into the tail vein. Results CXCR4 silenced with shRNA lead to a significant decrease in the capacity of the invasion and proliferation of melanoma cell in vitro. Moreover, melanoma cells with CXCR4 shRNA permanant transfection had much lower metastatic potential in nude mice than control cells and mock control cells in vivo. The expression of MMP-2 was down-regulated in shRNA group. Conclusion Silencing of CXCR4 by shRNA lead to significant decrease in melanoma cell invasion and proliferation capacity of melanoma in vitro. CXCR4 gene silencing mediated by shRNA significantly suppressed the growth of MV3 cell in vitro and effectively decrease the metastatic potential of lung and liver and brain. The block of SDF-1/CXCR4 pathway and suppression of MMP-2 may invlolve in its mechanism of anti-metastasis. |
| keywords:Melanoma Chemokin receptor CXCR4 Gene silencing Tumor metastasis |
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