摘要
目的:探讨趋化因子受体CXCR4表达水平对人肺癌细胞转移潜能的影响。方法:采用RT-PCR,FACS检测趋化因子受体CXCR4在肺癌细胞株95C,95D细胞的表达。构建CXCR4正义和反义真核表达质粒,用脂质体法转染至95C和95D细胞内,通过G418筛选出稳定转染株。通过趋化和趋化侵袭实验测定转染前后细胞对基质衍生因子(SDF-1α)的迁移能力,通过明胶酶谱法检测MMP-2活性,通过FACS检测细胞对血管内皮细胞的黏附能力。结果:CXCR4正义和反义真核表达质粒pcDNA3-X4和pcDNA3-ASX4能明显上调或下调肺癌细胞表面CXCR4的表达,上调95C细胞表面CXCR4的表达可以增强其对SDF-1α的趋化反应性、MMP-2活性及其对血管内皮细胞的黏附能力。相反,下调95D细胞表面CXCR4的表达抑制了其对SDF-1α的趋化反应性、MMP-2活性及其对血管内皮细胞的黏附能力。结论:上调或下调肺癌细胞表面CXCR4表达可以显著增强或抑制其转移潜能,提示CXCR4表达水平与肺癌细胞转移潜能有关。
Objective: To study the effect of the expression levels of Chemokine receptor CXCR4 on the metastatic potential of human lung cancer. Methods: CXCR4 expression was determined by Real-time PCR and FACS. The plasmid DNA containing CXCR4 coding gene or CXCR4 antisense nucleotides fragment was constructed and transfected into 95 C or 95D cells with Lipofectamine 2000 reagent respectively, and then the stable transfectants were screened by G418. Migratory responses to SDF-1 α were detected by chemotaxis and chemoinvasion assay, MMP-2 activity was determined with zy-mography, and the ability of adhesion to ECV-304 cells was analyzed by FACS. Results: The surface expression of CX-CR4 on lung cancer cells was significantly up or down regulated. Following up-regulation of CXCR4 expression on 95C cells, the migratory response to SDF-1 α, MMP-2 activity, and the ability of adhesion to ECV-304 cells were consequently enhanced. Conversely, when CXCR4 expression was down regulated on 95D cells, the migratory response to SDF-1 α, MMP-2 activity, and the ability of adhesion to ECV-304 cells were significantly impaired. Conclusion: Up or down regulation of CXCR4 expression enhanced or inhibited the metastatic potential of human lung cancer cells, which implied that CXCR4 expression was closely associated with the metastatic potential of human lung cancer cells.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2005年第1期46-51,共6页
Chinese Journal of Cancer Biotherapy
基金
国家教育部科学技术重点项目(03063)国家杰出青年科学基金(39925031)
关键词
CXCR4
肺癌细胞
转移潜能
CXCR4
human lung cancer cell
metastatic potential
